Recombinant Rat Agrin Protein Summary
Details of Functionality |
Measured by its ability to induce acetylcholine receptor clustering on myotubes differentiated from C2C12 mouse myoblast cells. Ferns, M.J. et al. (1993) Neuron 11:491. The ED50 for this effect is 2-6 ng/mL. Measured by its binding ability in a functional ELISA. When recombinant human LRP-4 is coated at 0.5 μg/mL, Recombinant Rat Agrin binds with an apparent Kd <3 nM. |
Source |
Spodoptera frugiperda, Sf 21 (baculovirus)-derived rat Agrin protein Ala1153-Pro1959 (Pro1788-Ser1798 del), with an N-terminal Met and 6-His tag |
Accession # |
|
N-terminal Sequence |
Met |
Protein/Peptide Type |
Recombinant Proteins |
Gene |
Agrn |
Purity |
>90%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining. |
Endotoxin Note |
<0.10 EU per 1 μg of the protein by the LAL method. |
Applications/Dilutions
Dilutions |
|
Theoretical MW |
90 kDa. Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors. |
SDS-PAGE |
100 kDa, reducing conditions |
Publications |
Read Publications using 550-AG in the following applications:
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Packaging, Storage & Formulations
Storage |
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.- 12 months from date of receipt, -20 to -70 °C as supplied.
- 1 month, 2 to 8 °C under sterile conditions after reconstitution.
- 3 months, -20 to -70 °C under sterile conditions after reconstitution.
|
Buffer |
Lyophilized from a 0.2 μm filtered solution in PBS with BSA as a carrier protein. |
Purity |
>90%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining. |
Reconstitution Instructions |
Reconstitute at 500 μg/mL in sterile PBS containing at least 0.1% human or bovine serum albumin. |
Notes
This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.
Alternate Names for Recombinant Rat Agrin Protein
Background
Agrin is a 400‑600 kDa heparan sulfate proteoglycan component of the extracellular matrix. The N‑terminal half of rat Agrin, which mediates ECM interactions, contains nine Kazal-type protease inhibitor domains, two Laminin EGF-like domains, and one SEA domain. The C-terminal half contains four EGF-like repeats and three Laminin globular G domains. Human Agrin also contains a Laminin‑binding N-terminal Agrin domain (NtA), and mouse and chick Agrin include the NtA domain only by the use of an alternate promoter. Additional isoforms are generated by alternate splicing at sites Y and Z in the C-terminal half of rat Agrin (known as A and B, respectively in chick). Agrin isoforms that contain an insert at site Z (Z+ forms) are known as neural Agrin and are selectively produced by motoneurons. Other isoforms are known as muscle Agrin and are additionally expressed in non-neuronal tissues, particularly in basement membranes of the lung and kidney (1-3). This recombinant protein consists of the C-terminal half of rat Agrin and contains a nine amino acid (aa) insert at the Z site. It shares 59%, 80%, and 94% aa sequence identity with comparable regions of chick, human, and mouse Agrin, respectively. The C-terminal half of Z- and Z+ Agrin binds to alpha -Dystroglycan and mediates adhesion between motoneurons and myotubes at the neuromuscular junction (NMJ) (4-6). In contrast, only Z+ Agrin is effective at inducing clustering of the postsynaptic Acetylcholine Receptor (AChR) and presynaptic motoneuron differentiation (7, 8). Agrin‑induced AChR clustering requires a myotube receptor complex that contains alpha -Dystroglycan, MuSK, and LRP4 (4, 9-11). Agrin exhibits many functions in addition to NMJ development. It is enriched in senile Alzheimer’s disease plaques where it binds the A beta (1-40) peptide and promotes amyloid fibril formation (12). It regulates neuronal excitability by binding and inhibiting the alpha 3 subunit of the neuronal Na/K ATPase (13). It functions as an epithelial cell attachment receptor for HIV-1 through interactions with the gp41 coat protein (14). During T cell activation, Agrin contributes to formation of the immunological synapse and regulates the threshold of T cell activation (15).
- Jury, E.C. and P.S. Kabouridis (2010) Arthritis Res. Ther. 12:205.
- Bezakova, G. and M.A. Ruegg (2003) Nat. Rev. Mol. Cell Biol. 4:295.
- Rupp, F. et al. (1991) Neuron 6:811.
- Gee, S.H. et al. (1994) Cell 77:675.
- Sugiyama, J. et al. (1994) Neuron 13:103.
- Gesemann, M. et al. (1998) J. Biol. Chem. 273:600.
- Burgess, R.W. et al. (1999) Neuron 23:33.
- Ferns, M.J. et al. (1993) Neuron 11:491.
- Glass, D.J. et al. (1996) Cell 85:513.
- Kim, N. et al. (2008) Cell 135:334.
- Zhang, B. et al. (2008) Neuron 60:285.
- Cotman, S.L. et al. (2000) Mol. Cell. Neurosci. 15:183.
- Hilgenberg, L.G.W. et al. (2006) Cell 125:359.
- Alfsen, A. et al. (2005) Mol. Biol. Cell 16:4267.
- Khan, A.A. et al. (2001) Science 292:1681.
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