Technical Data
A1059-60B
Agrin
Description:
Agrin is an essential extracellular matrix component which promotes clustering of nicotinic acetylcholine receptors (nAChRs) and other proteins during development at the neuromuscular junction (3, 5). Agrin, MuSK and Rapsyn are all essential components for AChR aggregation, through an unknown mechanism. The C-terminal region of agrin is released into the medium, interacts with receptors on the muscle surface and induces AChR aggregation. The central region contains two O-linked glycosylation sites and a domain homologous to domain III of laminin. The N-terminal region anchors agrin to the extracellular matrix via other basal membrane components. This region also contains a protease inhibitor domain and glycosaminoglycan attachment sites; increasing the predicted MW from 200kD to ~600kD. The diagram below indicates the domain structure and functional regions of agrin, as well as domains required for AChR aggregation and alpha-dystroglycan and heparin binding (5): Various agrin isoforms are generated by alternative splicing at the X, Y and Z sites and differ in the pressence or absence of small inserts. The isoforms can determine the biological activity of agrin and their expression in specific tissues and stages of development. (5). While no difference in functional activity has been detected between splicing variants at site X (2), insertion of a 4 amino acid (aa) peptide at site Y modestly increases agrins nAChR clustering activity (2,4). Insertion of an 8aa peptide at splicing site Z increases the clustering activity of soluble rat agrin 10,000 fold (2).

Applications:
Suitable for use in Immunohistochemistry. Other applications not tested.

Recommended Dilution:
Immunohistochemistry: 1:100
Optimal dilutions to be determined by the researcher.

Storage and Stability:
May be stored at 4C for short-term only. For long-term storage, store at -20C. Aliquots are stable for at least 12 months at -20C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.
TypeIsotypeCloneGrade
MabIgG14E5Affinity Purified
SizeStorageShippingSourceHost
50ug-20CBlue IceRatMouse
Concentration:
As reported
Immunogen:
Recombinant rat agrin, C-terminal construct
Purity:
Purified by Protein G affinity chromatography.
Form
Supplied as a liquid in PBS, pH 7.2, 0.09% azide, 50% glycerol.
Specificity:
Recognizes mouse and rat agrin. Epitope has been mapped near the 2nd laminin domain homology region and causes a marked reduction in the number of nicotinic acetylcholine receptor (nAChR) clusters. Identifies recombinant rat agrin on immunoblots.
Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.
1. Hoch, W., Campanelli, J.T., Harrison, S., and Scheller, R.H., EMBO J. 13: 28142821 (1994). 2. Ferns, M.J., Campanelli, J.T., Hoch, W., Scheller, R.H., and Hall, Z.; Neuron. 11: 491502 (1993). 3. Wallace, B.G.; J. Neurosci., 9: 12941302 (1989). 4. Tsim, K.W.K., Ruegg, M.A., Escher, G., Kroger, S. and McMahon, U.J. Neuron 8: 677689 (1992). 5. Hoch, W. Eur J Biochem 265: 110 (1999). 6. Groffen, A.J., et al.; Histochem Cytochem. 46: 1928 (1998). 7. Kh an, A.A., Bose, C., Yam, L.S., Soloski, M.J. and Rupp, F.; Science 292: 16811686 (2001).