Recombinant Mouse GPVI Protein, CF Summary
Product Specifications
When cross-linked with 10 µg/mL of Mouse Anti-polyHistidine Monoclonal Antibody (Catalog # MAB050), Recombinant Mouse GPVI binds to Collagen I (coated at 10 µg/mL, 100 µL/well) with an apparent KD <90 nM.
Optimal dilutions should be determined by each laboratory for each application.
Met1-Lys265, with a C-terminal 6-His tag
Analysis
Product Datasheets
Carrier Free
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.
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.
6758-GP
| Formulation | Lyophilized from a 0.2 μm filtered solution in PBS. |
| Reconstitution | Reconstitute at 400 μg/mL in 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.
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Reconstitution Calculator
Background: GPVI
Glycoprotein VI (GPVI) is a 63 kDa platelet/megakaryocyte-specific type I transmembrane glycoprotein of the immunoglobulin superfamily that is an important collagen receptor and initiator of platelet activation, aggregation and thrombus generation (1, 2). GPVI is also a secondary receptor required for platelet spreading on laminin (3). Mouse GPVI contains a 21 amino acid (aa) signal sequence, a 244 aa extracellular domain (ECD) that has two C‑type Ig‑like domains followed by a mucin‑like, presumably O‑glycosylated Ser‑Thr‑rich region, a 21 aa transmembrane (TM) domain and a 27 aa cytoplasmic tail. Mouse GPVI ECD shares 70%, 85%, 69% and 66% aa identity with human, rat, equine and canine GPVI ECD, respectively. A 296 aa mouse isoform lacking aa 224 ‑ 240 has been described (4). GPVI associates with the Fc receptor gamma ‑chain via charged aa in the TM domains of GPVI (arginine) and the FcR gamma (aspartic acid) (2). Collagen binding by the GPVI Ig‑like domains initiates signaling through the FcR gamma ITAM sequence (2). Dimerization of GPVI (2 CPVI monomers: FcR gamma dimer) and N‑glycosylation greatly enhances collagen binding (5, 6). Type I and III collagens are strong thrombus‑forming components in the vascular subendothelium and atherosclerotic plaques (7). GPVI initiates binding to fibrillar collagens under flow conditions, then activates integrin alpha 2 beta 1 which binds collagen more tightly (8). GPVI deficiencies cause only a mild bleeding tendency, in part because integrin alpha 2 beta 1 is able to minimally initiate collagen binding (8). Engagement of GPVI by collagens or other agonists, including GPVI antibodies, causes cleavage of the 57 kDa ECD by ADAM10, TACE/ADAM17, or other proteases, depleting surface GPVI (9, 10). In rheumatoid arthritis, collagen engagement of synovial platelet GPVI produces platelet microparticles that contribute to inflammation (11).
- Jandrot-Perrus, M. et al. (2000) Blood 96:1798.
- Moroi, M. and S. M. Jung (2004) Thromb. Res. 114:221.
- Inoue, O. et al. (2006) Blood 107:1405.
- GenBank protein accession #AAI45173.
- Horii, K. et al. (2006) Blood 108:936.
- Kunicki, T. J. et al. (2005) Blood 106:2744.
- Cosemans, J. M. et al. (2005) Atherosclerosis 181:19.
- Lecut, C. et al. (2005) Thromb. Haemost. 94:107.
- Stephens, G. et al. (2005) Blood 105:186.
- Bender, M. et al. (2010) Blood 116:3347.
- Boilard, E. et al. (2010) Science 327:580.
FAQs
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This protein datasheet indicates I need to use a cross-linking antibody, Catalog # MAB050, for biological activity. What is this antibody and is it really necessary?
The antibody is directed against a 6x histidine repeat and is recommended for use as a cross-linker of proteins with 6x his-tag. Crosslinking is often used for proteins that require receptor trimerization and can result greater biological activity. R&D Systems Quality Control tests the performance of these proteins in the presence of the cross-linking antibody. Therefore, it is necessary to use this antibody when trying to achieve the same level of specific activity described in the datasheet.
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