Specifications

Product Data
Applications	ELISA, IF, IHC, WB
Recommended Dilution	Polyclonal anti-GFP antibody is designed to detect GFP and its variants. This antibody can be used to detect GFP by ELISA (Sandwich or Capture) for the direct binding of antigen and recognizes wild type, recombinant and enhanced forms of GFP. Biotin conjugated polyclonal anti-GFP antibody used in a Sandwich ELISA is well suited to titrate GFP in solution using this antibody in combination with a monoclonal anti-GFP antibody (Cat.-No R1461P) using either form of the antibody as the Capture or Detection antibodies. However, use the monoclonal form only for the detection of wild type or recombinant GFP as this form does not sufficiently detect 'enhanced' GFP. The detection antibody is typically conjugated to biotin and subsequently reacted with streptavidin conjugated HRP (Cat.-No RA021HRP). Fluorochrome conjugated polyclonal anti-GFP antibody can be used to detect GFP by Immunofluorescence Microscopy in prokaryotic (E. coli) and eukaryotic (CHO cells) expression systems and can detect GFP containing inserts. Significant amplification of signal is achieved using fluorochrome conjugated polyclonal anti-GFP antibody relative to the fluorescence of GFP alone. For Immunoblotting use either alkaline phosphatase or peroxidase conjugated polyclonal anti-GFP antibody to detect GFP or GFP containing proteins on western blots. Recommended Dilutions: ELISA: 1/10,000-1/30,000. Immunofluorescence: 1/500. Western blot: 1/1,000-1/10,000. Immunohistochemistry: 1/200-1/1,000.
Host	Goat
Clonality	Polyclonal
Immunogen	Green Fluorescent Protein (GFP) fusion protein corresponding to the full length amino acid sequence (246 aa) derived from the jellyfish Aequorea victoria
Specificity	Detects wt GFP, rGFP and eGFP. Assay by Immunoelectrophoresis resulted in a single precipitin arc against anti-Goat serum and purified and partially purified Green Fluorescent Protein (Aequorea victoria) serum. No reaction was observed against Human, Mouse or Rat serum proteins.
Formulation	0.02M Potassium Phosphate, 0.15M Sodium Chloride, pH 7.2 State: Aff - Purified State: Liquid (sterile filtered) purified Ig fraction Stabilizer: None Preservative: 0.01% (w/v) Sodium Azide
Concentration	lot specific
Purification	Immunoaffinity Chromatography using Green Fluorescent Protein (Aequorea victoria) coupled to agarose beads followed by solid phase adsorption(s) to remove any unwanted reactivities
Conjugation	Unconjugated
Storage	Store undiluted at 2-8°C for one month or (in aliquots) at -20°C for longer. Avoid repeated freezing and thawing.
Stability	Shelf life: one year from despatch.
Database Link	UniProt ID P42212
Background	Green fluorescence protein (GFP) is a 27 kDa protein derived from the jellyfish Aequorea victoria, which emits green light (emission peak at a wavelenth of 509 nm) when excited by blue light (excitation peak at a wavelenth of 395 nm). Green Fluorescent Protein (GFP) has become an invaluable tool in cell biology research, since its intrinsic fluorescence can be visualized in living cells. GFP fluorescence is stable under fixation conditions and suitable for a variety of applications. GFP has been widely used as a reporter for gene expression, enabling researchers to visualize and localize GFP-tagged proteins within living cells without the need for chemical staining. Other applications of GFP include assessment of protein protein interactions through the yeast two hybrid system and measurement of distance between proteins through fluorescence energy transfer (FRET) protocols. GFP technnology has considerably contributed to a greater understanding of cellular physiology. YFP differs from GFP due to a mutation at T203Y; antibodies raised against full-length GFP should also detect YFP and other variants.
Synonyms	Green fluorescent protein, GFP-Tag
Reference Data