Recombinant Human GPVI Fc Chimera Protein, CF

Name: Recombinant Human GPVI Fc Chimera Protein, CF
Brand: R&D Systems
SKU: 10452-GP
Rating: 4 (1 reviews)

Catalog #: 10452-GP 1 Review Datasheet / COA / SDS

HEK293 Expressed

Catalog #	Availability	Size / Price	Qty
10452-GP-050

Recombinant Human GPVI Fc Chimera Protein Binding Activity

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All GPVI Proteins and Enzymes

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Summary Product Datasheets Carrier Free Data Images Reconstitution Calculator Background Related Research Areas

Recombinant Human GPVI Fc Chimera Protein, CF Summary

Product Specifications

Purity

>95%, 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.

Activity

Measured by its binding ability in a functional ELISA. When Bovine Collagen I is immobilized at 10 µg/mL (100 μL/well), Recombinant Human GPVI Fc Chimera (Catalog # 10452-GP) binds with an ED₅₀ of 0.04-0.36 μg/mL.

Source

Human embryonic kidney cell, HEK293-derived human GPVI protein

Human GPVI (Gln21-Lys267) Accession # Q9HCN6.4	IEGRMD	Human IgG₁ (Pro100-Lys330)
N-terminus		C-terminus

Accession #

Q9HCN6.4

N-terminal Sequence
Analysis

Gln21

Structure / Form

Disulfide-linked homodimer

Predicted Molecular Mass

54 kDa

SDS-PAGE

68-76 kDa, under reducing conditions

Product Datasheets

10452-GP

Product Datasheet

COA

SDS

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.

10452-GP

Formulation	Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose.
Reconstitution	Reconstitute at 500 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. 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.

Scientific Data

Binding Activity

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When Bovine Collagen I is immobilized at 10 μg/mL (100 μL/well), Recombinant Human GPVI Fc Chimera (10452-GP) binds with an ED₅₀ of 0.04-0.36 μg/mL.

SDS-PAGE

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2 μg/lane of Recombinant Human GPVI Fc Chimera Protein (Catalog # 10452-GP) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 68-80 kDa and 136-160 kDa, respectively.

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 thrombin generation (1, 2). GPVI is also a secondary receptor required for platelet spreading on laminin (3). Human GPVI contains a 20 amino acid (aa) signal sequence, a 247 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 51 aa cytoplasmic tail that contains calmodulin-binding and SH3 domains. Human GPVI ECD shows 69%, 65% and 70% aa identity with mouse, bovine and canine GPVI ECD, respectively. Two splice variants exist; one is 17 aa shorter in the ECD, while the other diverges at aa 260, creating an inactive monomeric and presumably secreted 681 aa protein (3).GPVI associates with the FcR gamma via charged amino acid 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 (2:2 with FcR gamma ) 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, probably because integrin alpha 2 beta 1 is able to minimally initiate collagen binding (8). Normal human GPVI concentration can vary widely and affect maximum thrombin generation (9). Engagement of GPVI by collagens or other agonists, including autoantibodies, causes calmodulin-regulated metalloproteinase cleavage of the 57 kDa ECD and depletes surface GPVI (10).

References