Rabbit TrueBlot®: Anti-Rabbit IgG HRP
Cat# 18-8816-31
Size : 50ul
Brand : Rockland Immunochemicals
Contact local distributor :
Phone : +1 850 650 7790
Background
Rabbit IgG TrueBlot® is a unique horseradish peroxidase conjugated Anti-Rabbit IgG monoclonal secondary antibody. Rabbit IgG TrueBlot® enables detection of immunoblotted target protein bands, without hindrance by interfering immunoprecipitating immunoglobulin heavy and light chains. It is easy to generate publication-quality IP/Western Blot data with Rabbit IgG TrueBlot®, simply substitute the conventional HRP Anti-Rabbit IgG blotting reagent with Rabbit IgG TrueBlot® and follow the prescribed protocol for sample preparation and immunoblotting. Rabbit IgG TrueBlot® is ideal for use in protocols involving immunoblotting of immunoprecipitated proteins. TrueBlot preferentially detects the non-reduced form of rabbit IgG over the reduced, SDS-denatured form of IgG. When the immunoprecipitate is fully reduced immediately prior to SDS-gel electrophoresis, reactivity of Rabbit IgG TrueBlot® with the 55 kDa heavy chains and the 23 kDa light chains of the immunoprecipitating antibody is minimized thereby eliminating interference by the heavy and light chains of the immunoprecipitating antibody in IP/Western blot applications. Applications include studies examining post-translational modification (e.g., phosphorylation or acetylation) or protein-protein interactions.
Specifications for Rabbit TrueBlot®: Anti-Rabbit IgG HRP
Product Details
Rabbit TrueBlot®: Anti-Rabbit IgG HRP - 18-8816-31
Anti-Rabbit IgG HRP, TrueBlot, HRP TrueBlot ULTRA, Peroxidase TrueBlot, TrueBlot for IP/WB, TrueBlot for immunoprecipitation, TrueBlot for western blotting
Mouse
Peroxidase (Horseradish) ULTRA
Monoclonal
IgG
Immunoprecipitation Kit
Target Details
Rabbit
Rabbit TrueBlot® Antibody Peroxidase Conjugate was prepared from tissue culture supernatant by Protein G affinity chromatography. Assay by immunoelectrophoresis resulted in a single precipitin arc against Anti-Rabbit Serum. Reactivity is observed against native Rabbit IgG by both Western blot and ELISA.
Application Details
ELISA, IP, WB
ChIP, FISH, IF - View References
Rabbit IgG HRP TrueBlot has been tested in ELISA, Western blot, and immunoprecipitation and may also be used for detection in immunoblotting assays that do not employ immunoprecipitation. Rabbit IgG TrueBlot® is provided as 1000X solution. To conserve reagent, we recommend incubating the blots from minigels in sealed bags (removing as much air as possible) with minimal volume (2-3 mLs). If used conservatively at 2.5 mLs/blot will yield enough reagent for 20 blots. Note that there are three key procedural considerations: 1. Protein A or G should not be used for the immunoprecipitation. Use of protein A or G beads with the rabbit TrueBlot will result in contaminating bands. For immunoprecipitation, Anti-Rat IgG beads, or Anti-Rabbit IgG beads should be used for rat or rabbit immunoprecipitating antibodies, respectively. 2. Immunoprecipitate should be completely reduced. 3. BLOTTO/Milk should be used as the blocking protein for the immunoblot. All recommended dilutions for listed applications are intended as an initial recommendation, specific conditions for each protein and antibody combination should be specifically optimized by the end user.
Formulation
Liquid (sterile filtered)
1.0 mg/mL by UV absorbance at 280 nm
0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2
Proclin is added as an antimicrobial agent.
0.1 mg/ml Bovine Serum Albumin (BSA) - IgG and Protease free, 50% (v/v) Glycerol
Shipping & Handling
Wet Ice
Store at -20°C. This product is guaranteed for 6 months upon receipt, when handled and stored as instructed.
Expiration date is six (6) months from date of receipt.
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Meyer SN, Scuoppo C, Vlasevska S, et al. (2019). Unique and Shared Epigenetic Programs of the CREBBP and EP300 Acetyltransferases in Germinal Center B Cells Reveal Targetable Dependencies in Lymphoma. Immunity.
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Ma X, Wang D, Li N, Gao P, Zhang M, Zhang Y. (2019). Hippo kinase NDR2 inhibits IL-17 signaling by promoting Smurf1-mediated MEKK2 ubiquitination and degradation. Mol Immunol.
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Pazos M, Peters K, Casanova M, et al. (2019). Z-ring membrane anchors associate with cell wall synthases to initiate bacterial cell division. Nat Commun.
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Tedja R, Roberts CM, Alvero AB, et al. (2019). Protein kinase Cα-mediated phosphorylation of Twist1 at Ser-144 prevents Twist1 ubiquitination and stabilizes it. J Biol Chem.
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Kumar S, Gu Y, Abudu YP, et al. (2019). Phosphorylation of Syntaxin 17 by TBK1 Controls Autophagy Initiation. Dev Cell.
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Alexander RA, Lot I, Enslen H. (2019). Methods to Characterize Protein Interactions with β-Arrestin In Cellulo. Methods Mol Biol.
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Mitrofanova A, Mallela SK, Ducasa GM, et al. (2019). SMPDL3b modulates insulin receptor signaling in diabetic kidney disease. Nat Commun.
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Najafov A, Mookhtiar AK, Luu HS, et al. (2019). TAM Kinases Promote Necroptosis by Regulating Oligomerization of MLKL. Mol Cell.
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Gentsch GE, Spruce T, Owens NDL, Smith JC. (2019). Maternal pluripotency factors initiate extensive chromatin remodelling to predefine first response to inductive signals. Nat Commun.
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Forteza R, Ahsan MK, Cartón-García F, Arango D, Ameen NA, Salas PJ. (2019). Glucocorticoids and myosin5b loss of function induce heightened PKA signaling in addition to membrane traffic defects. Mol Biol Cell.
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Le-Bel G, Cortez Ghio S, Guérin LP, Bisson F, Germain L, Guérin SL. (2019). Irradiated Human Fibroblasts as a Substitute Feeder Layer to Irradiated Mouse 3T3 for the Culture of Human Corneal Epithelial Cells: Impact on the Stability of the Transcription Factors Sp1 and NFI. Int J Mol Sci.
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Lipinski S, Petersen BS, Barann M, et al. (2019). Missense variants in NOX1 and p22phox in a case of very-early-onset inflammatory bowel disease are functionally linked to NOD2. Cold Spring Harb Mol Case Stud.
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Boonying W, Joselin A, Huang E, et al. (2019). Pink1 regulates FKBP5 interaction with AKT/PHLPP and protects neurons from neurotoxin stress induced by MPP. J Neurochem.
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Paoletti A, Allouch A, Caillet M, et al. (2019). HIV-1 Envelope Overcomes NLRP3-Mediated Inhibition of F-Actin Polymerization for Viral Entry. Cell Rep.
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Bhattacharya T et al. (2019). Prdm9 and meiotic cohesin proteins cooperatively promote DNA double-strand break formation in mammalian spermatocytes. Curr Biol.
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Moriya et al. (2018). PRDM14 directly interacts with heat shock proteins HSP90α and glucose-regulated protein 78. Cancer Science
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Hartmann et al. (2018). β-Secretase BACE1 Promotes Surface Expression and Function of Kv3.4 at Hippocampal Mossy Fiber Synapses. The Journal of Neuroscience
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Haque et al. (2018). Development of an efficient chromatin immunoprecipitation method to investigate protein-DNA interaction in oleaginous castor bean seeds. PLOS One
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Sekiba et al. (2018). DHX9 regulates production of hepatitis B virus-derived circular RNA and viral protein levels. Oncotarget
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Mitchell et al. (2018). IL1RAP potentiates multiple oncogenic signaling pathways in AML. Journal of Experimental Medicine
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Perraki et al. (2018). Phosphocode-dependent functional dichotomy of a common co-receptor in plant signalling. Nature
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Cheng et al. (2018). The long non-coding RNA NEAT1 is elevated in polyglutamine repeat expansion diseases and protects from disease gene-dependent toxicities. Human Molecular Genetics
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Menzies et al. (2018). The sterol-responsive RNF145 E3 ubiquitin ligase mediates the degradation of HMG-CoA reductase together with gp78 and Hrd1. Elife
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Kumar et al. (2018). Mechanism of Stx17 recruitment to autophagosomes via IRGM and mammalian Atg8 proteins. Journal of Cell Biology
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Peña-Philippides et al. (2018). Inhibition of MicroRNA-155 Supports Endothelial Tight Junction Integrity Following Oxygen-Glucose Deprivation. Journal of the American Heart Association
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D'Souza et al. (2018). Gankyrin Promotes Tumor-Suppressor Protein Degradation to Drive Hepatocyte Proliferation. Cellular and Molecular Gastroenterology and Hepatology
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Predescu et al. (2018). Epsin15 Homology Domains: Role in the Pathogenesis of Pulmonary Arterial Hypertension. Frontiers in Physiology
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Pazos et al. (2018). Z-ring membrane anchors associate with cell wall synthases to initiate bacterial cell division. Nature Communications
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Kumar A, Giri S, Shaha C. (2018). Sestrin2 facilitates glutamine-dependent transcription of PGC-1α and survival of liver cancer cells under glucose limitation. FEBS J.
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Dai L, Zhao T, Bisteau X, et al. (2018). Modulation of Protein-Interaction States through the Cell Cycle. Cell.
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Choi SW, Park HH, Kim S, et al. (2018). PELI1 Selectively Targets Kinase-Active RIP3 for Ubiquitylation-Dependent Proteasomal Degradation. Mol Cell.
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Figlewicz DP, Jay J, West CH, et al. (2018). Effect of dietary palmitic and stearic acids on sucrose motivation and hypothalamic and striatal cell signals in the rat. Am J Physiol Regul Integr Comp Physiol.
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Gong L, Pan X, Lim CB, de Polo A, Little JB, Yuan ZM. (2018). A functional interplay between Δ133p53 and ΔNp63 in promoting glycolytic metabolism to fuel cancer cell proliferation. Oncogene.
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Zhu J, Zhang Y, Fan F, Wu G, Xiao Z, Zhou H. (2018). Tumor necrosis factor-α-induced protein 8-like-2 is involved in the activation of macrophages by Astragalus polysaccharides in vitro. Mol Med Rep.
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Hayashi K, Taura M, Iwasaki A. (2018). The interaction between IKKα and LC3 promotes type I interferon production through the TLR9-containing LAPosome. Sci Signal.
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Paradis-Isler N, Boehm J. (2018). NMDA receptor-dependent dephosphorylation of serine 387 in Argonaute 2 increases its degradation and affects dendritic spine density and maturation. J Biol Chem.
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Kita Y, Katayama Y, Shiraishi T, et al. (2018). The Autism-Related Protein CHD8 Cooperates with C/EBPβ to Regulate Adipogenesis. Cell Rep.
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Parikh VN, Liu J, Shang C, et al. (2018). Apelin and APJ orchestrate complex tissue-specific control of cardiomyocyte hypertrophy and contractility in the hypertrophy-heart failure transition. Am J Physiol Heart Circ Physiol.
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Krach F, Batra R, Wheeler EC, et al. (2018). Transcriptome-pathology correlation identifies interplay between TDP-43 and the expression of its kinase CK1E in sporadic ALS. Acta Neuropathol.
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Tykesson E, Hassinen A, Zielinska K, et al. (2018). Dermatan sulfate epimerase 1 and dermatan 4-O-sulfotransferase 1 form complexes that generate long epimerized 4-O-sulfated blocks. J Biol Chem.
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Yao B, Li Y, Wang Z, et al. (2018). Active N6-Methyladenine Demethylation by DMAD Regulates Gene Expression by Coordinating with Polycomb Protein in Neurons. Mol Cell.
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Dallaire A, Frédérick PM, Simard MJ. (2018). Somatic and Germline MicroRNAs Form Distinct Silencing Complexes to Regulate Their Target mRNAs Differently. Dev Cell.
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George E. Gentsch et al. (2018). The role of maternal pioneer factors in predefining first zygotic responses to inductive signals. bioRxiv
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Lee JM, Han HS, Jung YS, Harris RA, Koo SH, Choi HS. (2018). The SMILE transcriptional corepressor inhibits cAMP response element-binding protein (CREB)-mediated transactivation of gluconeogenic genes. J Biol Chem.
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Hou W, Cruz-Cosme R, Wen F, et al. (2018). Expression of Human Cytomegalovirus IE1 Leads to Accumulation of Mono-SUMOylated PML That Is Protected from Degradation by Herpes Simplex Virus 1 ICP0. J Virol.
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Lahaye X, Gentili M, Silvin A, et al. (2018). NONO Detects the Nuclear HIV Capsid to Promote cGAS-Mediated Innate Immune Activation. Cell. 2018;175(2):488-501.e22. doi:10.1016/j.cell.2018.08.062 Cell.
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Zhang et al. (2017). Protein Phosphotyrosine Phosphatase 1B (PTP1B) in Calpain-dependent Feedback Regulation of Vascular Endothelial Growth Factor Receptor (VEGFR2) in Endothelial Cells: IMPLICATIONS IN VEGF-DEPENDENT ANGIOGENESIS AND DIABETIC WOUND HEALING. Journal of Biological Chemistry
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Rivera-Serrano et al. (2017). A Cytoplasmic RNA Virus Alters the Function of the Cell Splicing Protein SRSF2. Journal of Virology
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Mansilla et al. (2017). Interference of the complex between NCS-1 and Ric8a with phenothiazines regulates synaptic function and is an approach for fragile X syndrome. Proc. Natl. Acad. Sci. U.S.A
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Huang et al. (2017). Dual mechanisms regulate the nucleocytoplasmic localization of human DDX6. Scientific Reports
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Goralska et al. (2017). Vitreous Humor Changes Expression of Iron-Handling Proteins in Lens Epithelial Cells. Investigative Ophthalmology & Visual Science
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Nakashima et al. (2017). Novel small molecule inhibiting CDCP1-PKCδ pathway reduces tumor metastasis and proliferation. Cancer Science
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Zhai et al. (2017). NLRP1 promotes tumor growth by enhancing inflammasome activation and suppressing apoptosis in metastatic melanoma. Oncogene
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Tagawa et al. (2017). Gene and protein expression of a soluble form of CTLA-4 in a healthy dog. Journal of Veterinary Medical Science
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Hu et al. (2017). An FAK-YAP-mTOR Signaling Axis Regulates Stem Cell-Based Tissue Renewal in Mice. Cell Stem Cell
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Miyoshi et al. (2017). DOK7 gene therapy enhances motor activity and life span in ALS model mice. EMBO Molecular Medicine
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Lewis et al. (2017). RNA Binding Protein CUGBP1 Inhibits Liver Cancer in a Phosphorylation-Dependent Manner. Molecular and Cellular Biology
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Joachim et al. (2017). Centriolar Satellites Control GABARAP Ubiquitination and GABARAP-Mediated Autophagy. Current Biology
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Van Nostrand et al. (2017). Variation in single-nucleotide sensitivity of eCLIP derived from reverse transcription conditions. Methods
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Weng et al. (2017). POPX2 phosphatase regulates apoptosis through the TAK1-IKK-NF-κB pathway. Cell Death & Disease
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Nagata et al. (2017). The Cdc45/RecJ-like protein forms a complex with GINS and MCM, and is important for DNA replication in Thermococcus kodakarensis. Nucleic Acids Research
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Mukherjee et al. (2017). A TNF-p100 pathway subverts noncanonical NF-κB signaling in inflamed secondary lymphoid organs. The EMBO Journal
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Short et al. (2017). p120-Catenin is an obligate haploinsufficient tumor suppressor in intestinal neoplasia. Journal of Clinical Investigation
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Lehmann et al. (2017). Mechanistic Insights into Autoinhibition of the Oncogenic Chromatin Remodeler ALC1. Molecular Cell
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Lu et al. (2017). Cell cycle-dependent phosphorylation regulates RECQL4 pathway choice and ubiquitination in DNA double-strand break repair. Nature Communications
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Seong et al. (2017). Zinc finger protein ZPR9 functions as an activator of AMPK-related serine/threonine kinase MPK38/MELK involved in ASK1/TGF-β/p53 signaling pathways. Scientific Reports
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Prevost et al. (2017). The Legionella effector WipB is a translocated Ser/Thr phosphatase that targets the host lysosomal nutrient sensing machinery. Scientific Reports
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Nagata et al. (2017). Possible function of the second RecJ-like protein in stalled replication fork repair by interacting with Hef. Scientific Reports
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Torregrosa-Muñumer R, Forslund JME, Goffart S, et al. (2017). PrimPol is required for replication reinitiation after mtDNA damage. Proc Natl Acad Sci U S A.
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Saud K, Cánovas J, Lopez CI, et al. (2017). SFPQ associates to LSD1 and regulates the migration of newborn pyramidal neurons in the developing cerebral cortex. Int J Dev Neurosci.
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Hayatsu N, Miyao T, Tachibana M, et al. (2017). Analyses of a Mutant Foxp3 Allele Reveal BATF as a Critical Transcription Factor in the Differentiation and Accumulation of Tissue Regulatory T Cells. Immunity.
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Iwahori S, Umaña AC, VanDeusen HR, Kalejta RF. (2017). Human cytomegalovirus-encoded viral cyclin-dependent kinase (v-CDK) UL97 phosphorylates and inactivates the retinoblastoma protein-related p107 and p130 proteins. J Biol Chem.
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Schering L, Albrecht E, Komolka K, Kühn C, Maak S. (2017). Increased expression of thyroid hormone responsive protein (THRSP) is the result but not the cause of higher intramuscular fat content in cattle. Int J Biol Sci.
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Iwahori S, Kalejta RF. (2017). Phosphorylation of transcriptional regulators in the retinoblastoma protein pathway by UL97, the viral cyclin-dependent kinase encoded by human cytomegalovirus. Virology.
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Perez KJ, Martínez FP, Cosme-Cruz R, Perez-Crespo NM, Tang Q. Correction for Perez et al., (2017). A Short cis-Acting Motif in the M112-113 Promoter Region Is Essential for IE3 To Activate M112-113 Gene Expression and Is Important for Murine Cytomegalovirus Replication. J Virol.
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Gonzalez-Calero et al. (2016). Urinary Kininogen-1 and Retinol binding protein-4 respond to Acute Kidney Injury: predictors of patient prognosis? Scientific Reports
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Maisonneuve et al. (2016). Molecular Basis of the Interaction of the Human Protein Tyrosine Phosphatase Non-receptor Type 4 (PTPN4) with the Mitogen-activated Protein Kinase p38γ. Journal of Biological Chemistry
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Aly et al. (2016). RNA Exosome Complex Regulates Stability of the Hepatitis B Virus X-mRNA Transcript in a Non-stop-mediated (NSD) RNA Quality Control Mechanism. Journal of Biological Chemistry
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Peng et al. (2016). Thioredoxin reductase 1 suppresses adipocyte differentiation and insulin responsiveness. Scientific Reports
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Hwang et al. (2016). Leucine Carboxyl Methyltransferase 1 (LCMT-1) Methylates Protein Phosphatase 4 (PP4) and Protein Phosphatase 6 (PP6) and Differentially Regulates the Stable Formation of Different PP4 Holoenzymes. Journal of Biological Chemistry
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Weaver et al. (2016). BubR1 alterations that reinforce mitotic surveillance act against aneuploidy and cancer. Elife
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Kishikawa et al. (2016). Satellite RNAs promote pancreatic oncogenic processes via the dysfunction of YBX1. Nature Communications
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Kunii et al. (2016). Opposing roles for SNAP23 in secretion in exocrine and endocrine pancreatic cells. Journal of Cell Biology
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Strikoudis et al. (2016). Regulation of transcriptional elongation in pluripotency and cell differentiation by the PHD-finger protein Phf5a. Nature Cell Biology
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Arima et al. (2016). Basigin can be a therapeutic target to restore the retinal vascular barrier function in the mouse model of diabetic retinopathy. Scientific Reports
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Lopez CI, Saud KE, Aguilar R, et al. (2016). The chromatin modifying complex CoREST/LSD1 negatively regulates notch pathway during cerebral cortex development. Dev Neurobiol.
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Jung B, Padula D, Burtscher I, et al. (2016). Pitchfork and Gprasp2 Target Smoothened to the Primary Cilium for Hedgehog Pathway Activation. PLoS One.
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Banerjee R, Russo N, Liu M, et al. : (2016). TRIP13 promotes error-prone nonhomologous end joining and induces chemoresistance in head and neck cancer. Nat Commun.
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Mian YA, Zeleznik-Le NJ. (2016). The miR-17∼92 cluster contributes to MLL leukemia through the repression of MEIS1 competitor PKNOX1. Leuk Res.
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Faouzi M, Hague F, Geerts D, et al. (2016). Functional cooperation between KCa3.1 and TRPC1 channels in human breast cancer: Role in cell proliferation and patient prognosis. Oncotarget.
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Masser DR, Clark NW, Van Remmen H, Freeman WM. (2016). Loss of the antioxidant enzyme CuZnSOD (Sod1) mimics an age-related increase in absolute mitochondrial DNA copy number in the skeletal muscle. Age (Dordr).
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Davies ER, Kelly JF, Howarth PH, et al. (2016). Soluble ADAM33 initiates airway remodeling to promote susceptibility for allergic asthma in early life. JCI Insight.
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Liu et al. (2015). Sequential actions of SIRT1-RELB-SIRT3 coordinate nuclear-mitochondrial communication during immunometabolic adaptation to acute inflammation and sepsis. Journal of Biological Chemistry
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Chung et al. (2015). Regulation of C-X-C chemokine gene expression by keratin 17 and hnRNP K in skin tumor keratinocytes. Journal of Cell Biology
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Kumar et al. (2015). MEI4 – a central player in the regulation of meiotic DNA double-strand break formation in the mouse. Journal of Cell Science
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Wani et al. (2015). Maturation of the proteasome core particle induces an affinity switch that controls regulatory particle association. Nature Communications
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Dagvadorj et al. (2015). Lipopolysaccharide Induces Alveolar Macrophage Necrosis via CD14 and the P2X7 Receptor Leading to Interleukin-1α Release. Immunity
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Schwartz et al. (2015). Eosinophil-specific deletion of IκBα in mice reveals a critical role of NF-κB-induced Bcl-xL for inhibition of apoptosis. Blood
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Greenfeld et al. (2015). TRAF1 Coordinates Polyubiquitin Signaling to Enhance Epstein-Barr Virus LMP1-Mediated Growth and Survival Pathway Activation. PLOS Pathogens
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Ibeawuchi et al. (2015). Hypertension-causing Mutations in Cullin3 Protein Impair RhoA Protein Ubiquitination and Augment the Association with Substrate Adaptors. Journal of Biological Chemistry
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Heneghan et al. (2015). BH3 domain-independent apolipoprotein L1 toxicity rescued by BCL2 prosurvival proteins. American Journal of Physiology-Cell Physiology
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Vuong et al. (2015). Differential Effects of Hepatocyte Nuclear Factor 4α Isoforms on Tumor Growth and T-Cell Factor 4/AP-1 Interactions in Human Colorectal Cancer Cells. Molecular and Cellular Biology
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Conde-Perez et al. (2015). A caveolin-dependent and PI3K/AKT-independent role of PTEN in β-catenin transcriptional activity. Nature Communications
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Schumacher et al. (2015). Shedding of Endogenous Interleukin-6 Receptor (IL-6R) Is Governed by A Disintegrin and Metalloproteinase (ADAM) Proteases while a Full-length IL-6R Isoform Localizes to Circulating Microvesicles. Journal of Biological Chemistry
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Sugawara S, Ito T, Sato S, et al. (2015). Generation of aminoterminally truncated, stable types of bioactive bovine and porcine fibroblast growth factor 4 in Escherichia coli. Biotechnol Appl Biochem.
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Marsh NM, Wareham A, White BG, Miskiewicz EI, Landry J, MacPhee DJ. (2015). HSPB8 and the Cochaperone BAG3 Are Highly Expressed During the Synthetic Phase of Rat Myometrium Programming During Pregnancy. Biol Reprod.
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Cauchy P, James SR, Zacarias-Cabeza J, et al. (2015). Chronic FLT3-ITD Signaling in Acute Myeloid Leukemia Is Connected to a Specific Chromatin Signature. Cell Rep.
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Cordier F, Chaffotte A, Wolff N. (2015). Quantitative and dynamic analysis of PTEN phosphorylation by NMR. Methods.
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Albrecht E, Kuzinski J, Komolka K, Gotoh T, Maak S. (2015). Localization and abundance of early markers of fat cell differentiation in the skeletal muscle of cattle during growth--are DLK1-positive cells the origin of marbling flecks? Meat Sci.
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Liang G, Liu G, Kitamura K, et al. (2015). TGF-β suppression of HBV RNA through AID-dependent recruitment of an RNA exosome complex. PLoS Pathog.
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Li H, Mohamed AA, Sharad S, et al. (2015). Silencing of PMEPA1 accelerates the growth of prostate cancer cells through AR, NEDD4 and PTEN. Oncotarget.
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Iwahori S, Hakki M, Chou S, Kalejta RF. (2015). Molecular Determinants for the Inactivation of the Retinoblastoma Tumor Suppressor by the Viral Cyclin-dependent Kinase UL97. J Biol Chem.
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Nishimura T. (2015). Role of extracellular matrix in development of skeletal muscle and postmortem aging of meat. Meat Sci.
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Heier C, Radner FP, Moustafa T, et al. (2015). G0/G1 Switch Gene 2 Regulates Cardiac Lipolysis. J Biol Chem.
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Zheng Q, Hou J, Zhou Y, Yang Y, Xie B, Cao X. (2015). Siglec1 suppresses antiviral innate immune response by inducing TBK1 degradation via the ubiquitin ligase TRIM27. Cell Res.
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Yousif AS et al. (2014). Differential regulation of S-region hypermutation and class switch recombination by noncanonical functions of uracil DNA glycosylase. Proc Natl Acad Sci USA
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Dusonchet J et al. (2014). A Parkinson's disease gene regulatory network identifies the signaling protein RGS2 as a modulator of LRRK2 activity and neuronal toxicity. Hum Mol Genet.
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Ogino H, Ishino S, Haugland GT, Birkeland NK, Kohda D, Ishino Y. (2014). Activation of the MCM helicase from the thermophilic archaeon, Thermoplasma acidophilum by interactions with GINS and Cdc6-2. Extremophiles.
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Amigo-Jiménez I, Bailón E, Ugarte-Berzal E, Aguilera-Montilla N, García-Marco JA, García-Pardo A. (2014). Matrix metalloproteinase-9 is involved in chronic lymphocytic leukemia cell response to fludarabine and arsenic trioxide. PLoS One.
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Puglisi R, Maccari I, Pipolo S, Mangia F, Boitani C. (2014). The nuclear form of glutathione peroxidase 4 colocalizes and directly interacts with protamines in the nuclear matrix during mouse sperm chromatin assembly. Spermatogenesis.
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Komolka K, Albrecht E, Schering L, Brenmoehl J, Hoeflich A, Maak S. (2014). Locus characterization and gene expression of bovine FNDC5: is the myokine irisin relevant in cattle? PLoS One.
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Totsuka N, Kim YG, Kanemaru K, et al. (2014). Toll-like receptor 4 and MAIR-II/CLM-4/LMIR2 immunoreceptor regulate VLA-4-mediated inflammatory monocyte migration. Nat Commun.
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Jerman S, Ward HH, Lee R, et al. (2014). OFD1 and flotillins are integral components of a ciliary signaling protein complex organized by polycystins in renal epithelia and odontoblasts. PLoS One.
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Baetz NW, Goldenring JR. (2014). Distinct patterns of phosphatidylserine localization within the Rab11a-containing recycling system. Cell Logist.
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Dannenberger D, Nuernberg K, Nuernberg G, Priepke A. (2014). Impact of dietary protein level and source of polyunsaturated fatty acids on lipid metabolism-related protein expression and fatty acid concentrations in porcine tissues. J Agric Food Chem.
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Gong H et al. (2013). Lipoprotein lipase (LPL) is associated with neurite pathology and its levels are markedly reduced in the dentate gyrus of Alzheimer's disease brains. The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society
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Baetz NW, Goldenring JR. (2013). Rab11-family interacting proteins define spatially and temporally distinct regions within the dynamic Rab11a-dependent recycling system. Mol Biol Cell.
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Applications
WB, IB, PCA PubMed
Gao Y, Koppen A, Rakhshandehroo M, et al. (2013). Early adipogenesis is regulated through USP7-mediated deubiquitination of the histone acetyltransferase TIP60. Nat Commun.
Applications
IP, Co-IP PubMed
Nelersa CM et al. (2012). High-content analysis of proapoptotic EphA4 dependence receptor functions using small-molecule libraries. J Biolmol Screen
Applications
WB, IB, PCA PubMed
Martínez FP, Tang Q. (2012). Leucine zipper domain is required for Kaposi sarcoma-associated herpesvirus (KSHV) K-bZIP protein to interact with histone deacetylase and is important for KSHV replication. J Biol Chem.
Applications
WB, IB, PCA PubMed
Albrecht E, Komolka K, Kuzinski J, Maak S. (2012). Agouti revisited: transcript quantification of the ASIP gene in bovine tissues related to protein expression and localization. PLoS One.
Applications
WB, IB, PCA PubMed
Medici D, Shore EM, Lounev VY, Kaplan FS, Kalluri R, Olsen BR. (2011). Conversion of vascular endothelial cells into multipotent stem-like cells. Nat Med.
Applications
IP, Co-IP PubMed
Vuletic S, Dong W, Wolfbauer G, Tang C, Albers JJ. (2011). PLTP regulates STAT3 and NFκB in differentiated THP1 cells and human monocyte-derived macrophages. Biochim Biophys Acta.
Applications
WB, IB, PCA PubMed
Cosme-Cruz R, Martínez FP, Perez KJ, Tang Q. (2011). H2B homology region of major immediate-early protein 1 is essential for murine cytomegalovirus to disrupt nuclear domain 10, but is not important for viral replication in cell culture. J Gen Virol.
Applications
WB, IB, PCA PubMed
Rzeczkowski K, Beuerlein K, Müller H, et al. (2011). c-Jun N-terminal kinase phosphorylates DCP1a to control formation of P bodies. J Cell Biol.
Applications
WB, IB, PCA PubMed
Chanda D, Xie YB, Choi HS. (2010). Transcriptional corepressor SHP recruits SIRT1 histone deacetylase to inhibit LRH-1 transactivation. Nucleic Acids Res.
Applications
WB, IB, PCA PubMed
Xie YB, Nedumaran B, Choi HS. (2009). Molecular characterization of SMILE as a novel corepressor of nuclear receptors. Nucleic Acids Res.
Applications
WB, IB, PCA PubMed
Xie YB, Park JH, Kim DK, et al. (2009). Transcriptional corepressor SMILE recruits SIRT1 to inhibit nuclear receptor estrogen receptor-related receptor gamma transactivation. J Biol Chem.
Applications
WB, IB, PCA; IP, Co-IP PubMed
Mitsuhashi S et al. (2008). Tautomycetin suppresses the TNFα/NF-κB pathway via inhibition of IKK activation. Int J Oncol
Applications
IP, Co-IP PubMed
Wolter S, Doerrie A, Weber A, et al. (2008). c-Jun controls histone modifications, NF-kappaB recruitment, and RNA polymerase II function to activate the ccl2 gene. Mol Cell Biol. 2008;28(13):4407-4423. doi:10.1128/MCB.00535-07 Mol Cell Biol.
Applications
WB, IB, PCA PubMed
Cozzone D, Fröjdö S, Disse E, et al. (2008). Isoform-specific defects of insulin stimulation of Akt/protein kinase B (PKB) in skeletal muscle cells from type 2 diabetic patients. Diabetologia.
Applications
WB, IB, PCA PubMed
Li H, Xu LL, Masuda K, et al. (2008). A feedback loop between the androgen receptor and a NEDD4-binding protein, PMEPA1, in prostate cancer cells. J Biol Chem.
Applications
WB, IB, PCA PubMed
Sun J, Lin X. (2008). Beta-arrestin 2 is required for lysophosphatidic acid-induced NF-kappaB activation. Proc Natl Acad Sci U S A.
Applications
WB, IB, PCA PubMed
Nelson RF, Glenn KA, Zhang Y, et al. (2007). Selective cochlear degeneration in mice lacking the F-box protein, Fbx2, a glycoprotein-specific ubiquitin ligase subunit. J Neurosci.
Applications
WB, IB, PCA PubMed
Woo SY, Kim DH, Jun CB, et al. (2007). PRR5, a novel component of mTOR complex 2, regulates platelet-derived growth factor receptor beta expression and signaling. J Biol Chem.
Applications
IP, Co-IP PubMed
Zhang AS, West AP Jr, Wyman AE, Bjorkman PJ, Enns CA. (2005). Interaction of hemojuvelin with neogenin results in iron accumulation in human embryonic kidney 293 cells. J Biol Chem.
Applications
WB, IB, PCA PubMed
Frequently Asked Questions (13)
What is Rockland’s TrueBlot® product line?
Rockland’s TrueBlot® product line is designed to solve common experimental problems when performing immunoprecipitation/co-immunoprecipitation experiments and Western blots of immunoprecipitated samples (IP–Western blot). The product line consists of TrueBlot® monoclonal secondary antibodies and TrueBlot® IP beads.
What is the advantage of TrueBlot® secondaries over regular secondary antibodies?
TrueBlot® antibodies are specific to the whole IgG molecule and will not bind heavy or light chains. This is useful for binding target proteins with an expected MW near 25 kDa (light chains) and 50 kDa (heavy chains).
Why does it appear that the TrueBlot® antibody is binding heavy/light chains in my sample?
The sample must be fully reduced to eliminate cross-reactivity with heavy and light chains. Any reactivity to heavy and light chains could be due to incomplete reduction of your sample. Please be sure to optimize reducing conditions for your sample type.
How can I ensure my samples are fully reduced?
Samples can be fully reduced by heating at 90-100°C for 10 minutes in sample buffer containing reducing agent (for example, DTT to a final concentration of 50 mM or add β-Mercaptoethanol to a final concentration of 2% (v/v)).
With which species/isotypes are TrueBlot® secondary antibodies reactive?
TrueBlot® secondary antibodies are reactive with the IgG of their respective species. For example, if using a primary antibody raised in mouse (mouse host), use TrueBlot® anti-mouse Ig secondary antibodies for detection.
Do TrueBlot® secondary antibodies detect IgM?
TrueBlot® secondary antibodies have not been shown to detect IgM.
Do I need to preclear my lysate before the immunoprecipitation step?
Samples that have many other proteins present (such as lysates) may require preclearing to prevent interference in later IP and WB procedures. Recombinant protein samples require pre-clearing more often than serum samples.
How can I enrich/concentrate my lysate for a low expression of a protein of interest?
The immunoprecipitation procedure can be repeated several times to yield a more concentrated protein solution.
What is the recommended lysis buffer for TrueBlot® products?
The optimal lysis buffer will depend on your sample type. See protocol for buffer recommendations. Generally, 1X RIPA is used to lyse samples.
My target protein has the same MW as a heavy/light chain. How can I be sure that the band is the target protein and not a heavy/light chain?
Be sure to include a positive control for the primary antibody in your experiment and check that the sample is fully reduced.
What is the average size of TrueBlot® magnetic/agarose beads?
The beads are roughly 0.5 μm in diameter.
Why can’t I use Protein A or Protein G beads for the IP step in IP–Western blot when using rabbit species?
Using Protein A or Protein G beads with rabbit species results in contaminated bands. For best results when using rabbit species, use Rockland’s TrueBlot® Rabbit Ig IP beads, which are available in either a magnetic or agarose format. Please see individual TrueBlot® Rabbit product pages for additional details.
Can I use Protein A or Protein G beads for the IP step in IP–Western blot when using mouse, goat, or sheep species?
Yes. If you are using Protein A or Protein G beads with mouse, goat, or sheep species, we recommend the following: Determine the compatibility of Protein A or Protein G with your species and subtype, do not use excessive amounts of slurry, and choose the appropriate elution conditions. In some instances, harsher elution conditions can cause stripping of the subunits of Protein A or Protein G from the bead support and result in non-specific bands. For best results, we recommend using TrueBlot® Ig IP beads, which are available in either agarose or magnetic format.
Related Protocols
Adherent Cell Lysis Protocol
Bifunctional Cross-Linking Protocol
Chromatin Immunoprecipitation (ChIP) Protocol
ELISA Protocol
Fluorescent Western Blotting Protocol
Histone Immunoblotting Protocol
Immunocytochemistry (ICC) Protocol
Immunofluorescence (IF) Protocol
Immunoprecipitation (IP) Protocol
In-Cell Western (ICW) Protocol
IP-WB with TrueBlot® Protocol
Multi-Lysate Western Blotting Protocol
Nuclear & Cytoplasmic Extract Protocol
Nucleosome ChIP Protocol
Re-ChIP Protocol
Sandwich ELISA Protocol for Collagen
Suspension Cultured Cell Lysis Protocol
Western Blotting (WB) Protocol
Certificate of Analysis Lookup
Disclaimer
This product is for research use only and is not intended for therapeutic or diagnostic applications. Please contact a technical service representative for more information. All products of animal origin manufactured by Rockland Immunochemicals are derived from starting materials of North American origin. Collection was performed in United States Department of Agriculture (USDA) inspected facilities and all materials have been inspected and certified to be free of disease and suitable for exportation. All properties listed are typical characteristics and are not specifications. All suggestions and data are offered in good faith but without guarantee as conditions and methods of use of our products are beyond our control. All claims must be made within 30 days following the date of delivery. The prospective user must determine the suitability of our materials before adopting them on a commercial scale. Suggested uses of our products are not recommendations to use our products in violation of any patent or as a license under any patent of Rockland Immunochemicals, Inc. If you require a commercial license to use this material and do not have one, then return this material, unopened to: Rockland Inc., P.O. BOX 5199, Limerick, Pennsylvania, USA.
Related products to: Rabbit TrueBlot®: Anti-Rabbit IgG HRP
Fluorescent TrueBlot®: Anti-Rabbit IgG DyLight™ 800 18-4516-32 Fluorescent TrueBlot®: Anti-Mouse Ig DyLight™ 680 18-4417-32 Fluorescent TrueBlot®: Anti-Rabbit IgG DyLight™ 680 18-4416-32 Fluorescent TrueBlot®: Anti-Rabbit IgG Fluorescein 18-0216-32 Fluorescent TrueBlot®: Anti-Mouse Ig DyLight™ 800 18-4517-32 Related Resources
Selecting TrueBlot® Products
Literature
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IP-WB with TrueBlot® Protocol
Protocols
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TrueBlot®: Publication Quality IP and Western Blot Images
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