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Recombinant Rat Agrin Protein

R&D Systems | Catalog # 550-AG

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Key Product Details

  • R&D Systems Sf 21 (baculovirus)-derived Recombinant Rat Agrin Protein (550-AG)
  • Quality control testing to verify active proteins with lot specific assays by in-house scientists
  • All R&D Systems proteins are covered with a 100% guarantee

Source

Sf 21 (baculovirus)

Accession Number

P25304.2

Applications

Bioactivity
View all Agrin Proteins and Enzymes »

Product Specifications

Source

Spodoptera frugiperda, Sf 21 (baculovirus)-derived rat Agrin protein
Ala1153-Pro1959 (Pro1788-Ser1798 del), with an N-terminal Met and 6-His tag

Purity

>90%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.

Endotoxin Level

<0.10 EU per 1 μg of the protein by the LAL method.

N-terminal Sequence Analysis

Met

Predicted Molecular Mass

90 kDa

SDS-PAGE

100 kDa, reducing conditions

Activity

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.

Reviewed Applications

Read 1 review rated 5 using 550-AG in the following applications:

Formulation, Preparation, and Storage

Carrier Free
What does CF mean?

CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.

What formulation is right for me?

In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.

Carrier: 550-AG
Formulation Lyophilized from a 0.2 μm filtered solution in PBS with BSA as a carrier protein.
Reconstitution Reconstitute at 500 μg/mL in sterile PBS containing at least 0.1% human or bovine serum albumin.
Shipping The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & 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.
Carrier Free: 550-AG/CF
Formulation Lyophilized from a 0.2 μm filtered solution in PBS.
Reconstitution Reconstitute at 100 μg/mL in sterile PBS.
Shipping The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & 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.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: Agrin

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).

References

  1. Jury, E.C. and P.S. Kabouridis (2010) Arthritis Res. Ther. 12:205.
  2. Bezakova, G. and M.A. Ruegg (2003) Nat. Rev. Mol. Cell Biol. 4:295.
  3. Rupp, F. et al. (1991) Neuron 6:811.
  4. Gee, S.H. et al. (1994) Cell 77:675.
  5. Sugiyama, J. et al. (1994) Neuron 13:103.
  6. Gesemann, M. et al. (1998) J. Biol. Chem. 273:600.
  7. Burgess, R.W. et al. (1999) Neuron 23:33.
  8. Ferns, M.J. et al. (1993) Neuron 11:491.
  9. Glass, D.J. et al. (1996) Cell 85:513.
  10. Kim, N. et al. (2008) Cell 135:334.
  11. Zhang, B. et al. (2008) Neuron 60:285.
  12. Cotman, S.L. et al. (2000) Mol. Cell. Neurosci. 15:183.
  13. Hilgenberg, L.G.W. et al. (2006) Cell 125:359.
  14. Alfsen, A. et al. (2005) Mol. Biol. Cell 16:4267.
  15. Khan, A.A. et al. (2001) Science 292:1681.

Alternate Names

AGRN

Entrez Gene IDs

375790 (Human); 11603 (Mouse); 25592 (Rat)

Gene Symbol

AGRN

UniProt

P25304.2

Additional Agrin Products

Product Documents for Recombinant Rat Agrin Protein

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

Download SDS 550-AG (with carrier)
Download SDS 550-AG/CF (carrier free)

Product Specific Notices for Recombinant Rat Agrin Protein

For research use only

Citations for Recombinant Rat Agrin Protein

Customer Reviews for Recombinant Rat Agrin Protein (1)

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  • Recombinant Rat Agrin Protein
    Name: Mia Huang
    Application: In vitro bioactivity in cell culture
    Verified Customer | Posted 05/23/2017
    Shown here are differentiated C2C12 mouse skeletal muscle myotubes treated with or without agrin (5 nM) in DMEM (1 hr, 37C). Subsequent staining with fluorophore-labeled-alpha bungarotoxin revealed robust AChR clusters (green) in agrin-treated myotubes.
    Recombinant Rat Agrin Protein 550-AG

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FAQs for Recombinant Rat Agrin Protein

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  • Q: What is the isoform of 550-AG?

    A: The source sequence is Ala1153-Pro1959 (Pro1788-Ser1798 del), Accession # P25304.2, with an N-terminal Met and 6-His tag.  This is Isoform 4 in the NCBI database.

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