Rat Agrn (Agrin) Sandwich ELISA Kit

Storage Instruction:	If unopened the kit may be stored at 2-8°C for up to 1 month. If the kit will not be used within 1 month, store the components separately, according to the component table in the manual.
Assay Time:	3.5h
Detection:	Colormetric

Gene Symbol:	Agrn
Gene ID:	25592
Uniprot ID:	AGRIN_RAT
Specificity:	This kit recognizes Rat AGRN in samples. No significant cross-reactivity or interference between Rat AGRN and analogues was observed.
Sample Type:	Serum, plasma and other biological fluids

Post Translational Modifications	Contains heparan and chondroitin sulfate chains and alpha-dystroglycan as well as N-linked and O-linked oligosaccharides. Glycosaminoglycans (GAGs), present in the agrin N-terminal 110 kDa fragment, are required for induction of filopodia in hippocampal neurons. The first cluster (Gly/Ser-rich) for GAG attachment contains heparan sulfate (HS) chains and the second cluster (Ser/Thr-rich), contains chondroitin sulfate (CS) chains. Heparin and heparin sulfate binding in the G3 doamin is independent of calcium ions. Binds heparin with a stoichiometry of 2:1. Binds sialic acid with a stoichiometry of 1:1 and binding requires calcium ions. At synaptic junctions, cleaved at two conserved sites, alpha and beta, by neurotrypsin. Cleavage at the alpha-site produces the agrin N-terminal 110-kDa subunit and the agrin C-terminal 110-kDa subunit. Further cleavage of agrin C-terminal 110-kDa subunit at the beta site produces the C-terminal fragments, agrin C-terminal 90 kDa fragment and agrin C-terminal 22 kDa fragment. Excessive cleavage at the beta-site releases large amounts of the agrin C-terminal 22 kDa fragment leading to destabilization at the neuromuscular junction (NMJ).
Function	Isoform 1: Heparan sulfate basal lamina glycoprotein that plays a central role in the formation and the maintenance of the neuromuscular junction (NMJ) and directs key events in postsynaptic differentiation. This neuron-specific (z+) isoform is a component of the AGRN-LRP4 receptor complex that induces the phosphorylation and activation of MUSK. The activation of MUSK in myotubes induces the formation of NMJ by regulating different processes including the transcription of specific genes and the clustering of AChR in the postsynaptic membrane. Calcium ions are required for maximal AChR clustering. AGRN function in neurons is highly regulated by alternative splicing, glycan binding and proteolytic processing. Modulates calcium ion homeostasis in neurons, specifically by inducing an increase in cytoplasmic calcium ions. Functions differentially in the central nervous system (CNS) by inhibiting the alpha(3)-subtype of Na+/K+-ATPase and evoking depolarization at CNS synapses. This transmembrane agrin (TM-agrin) isoform, the predominate form in neurons of the brain, induces dendritic filopodia and synapse formation in mature hippocampal neurons in large part due to the attached glycosaminoglycan chains and the action of Rho-family GTPases. Isoform 1, isoform 4, isoform 5 and isoform 6: neuron-specific (z+) isoforms that contain C-terminal insertions of 8-19 AA are potent activators of AChR clustering. Isoform 5, agrin (z+8), containing the 8-AA insert, forms a receptor complex in myotubules containing the neuronal AGRN, the muscle-specific kinase MUSK and LRP4, a member of the LDL receptor family. The splicing factors, NOVA1 and NOVA2, regulate AGRN splicing and production of the 'z' isoforms. Agrin N-terminal 110 kDa subunit: Is involved in regulation of neurite outgrowth probably due to the presence of the glycosaminoglcan (GAG) side chains of heparan and chondroitin sulfate attached to the Ser/Thr- and Gly/Ser-rich regions. Also involved in modulation of growth factor signaling. Agrin C-terminal 22 kDa fragment: This released fragment is important for agrin signaling and to exert a maximal dendritic filopodia-inducing effect. All 'z' splice variants (z+) of this fragment also show an increase in the number of filopodia.
Protein Name	Agrin Cleaved Into - Agrin N-Terminal 110 Kda Subunit - Agrin C-Terminal 110 Kda Subunit - Agrin C-Terminal 90 Kda Fragment C90 - Agrin C-Terminal 22 Kda Fragment C22
Database Links	Reactome: R-RNO-1971475 Reactome: -RNO-2022928 Reactome: -RNO-2024096 Reactome: -RNO-3000178 Reactome: -RNO-975634
Cellular Localisation	Synapse Cell Membrane Single-Pass Type Ii Membrane Protein
Alternative ELISA Names	Agrin Cleaved Into - Agrin N-Terminal 110 Kda Subunit - Agrin C-Terminal 110 Kda Subunit - Agrin C-Terminal 90 Kda Fragment ELISA kit C90 - Agrin C-Terminal 22 Kda Fragment ELISA kit C22 ELISA kit Agrn ELISA kit Agrin ELISA kit
Specificity	This kit recognizes Rat AGRN in samples. No significant cross-reactivity or interference between Rat AGRN and analogues was observed.
Reproducibility	Both intra-CV and inter-CV are

Information sourced from Uniprot.org

Item	Specifications	Storage
Micro ELISA Plate (Dismountable)	96T: 8 wells ×12 strips strips	-20℃, 6 months
Reference Standard	96T: 2 vials 48T: 1 vial	-20℃, 6 months
Concentrated Biotinylated Detection Ab (100×)	96T: 1 vial, 120 μL 60 μL	-20℃, 6 months
Concentrated HRP Conjugate (100×)	96T: 1 vial, 120 μL 60 μL	-20℃ (Protect from light), 6 months
Reference Standard & Sample Diluent	1 vial, 20 mL	2-8°C, 6 months
Biotinylated Detection Ab Diluent	1 vial, 14 mL	2-8°C, 6 months
HRP Conjugate Diluent	1 vial, 14 mL	2-8°C, 6 months
Concentrated Wash Buffer (25×)	1 vial, 30 mL	2-8°C, 6 months
Substrate Reagent	1 vial, 10 mL	2-8℃ (Protect from light)
Stop Solution	1 vial, 10 mL	2-8°C
Plate Sealer	5 pieces
Manual	1 copy
Certificate of Analysis	1 copy

Sample Type	Range (%)	Average Recovery (%)
Serum(n=8)	93-107	100
EDTA plasma (n=8)	87-98	93
Cell culture media (n=8)	88-104	95

	Intra-assay Precision	Intra-assay Precision	Intra-assay Precision	Inter-assay Precision	Inter-assay Precision	Inter-assay Precision
Sample	1.00	2.00	3.00	1.00	2.00	3.00
n	20.00	20.00	20.00	20.00	20.00	20.00
Mean (pg/mL)	104.05	257.27	686.98	110.86	277.63	701.40
Standard deviation	6.54	10.63	31.19	7.74	14.85	24.27
CV (%)	6.29	4.13	4.54	6.98	5.35	3.46

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