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Biotinylated Human VEGF165 Protein, Avi Tag (Avitag™)  pdf  pdf  pdf


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Synonym

RP1-261G23.1,MGC70609,MVCD1,VEGFA,VPF

Source

Biotinylated Human VEGF165, His Tag (VE5-H82Q0) is expressed from human 293 cells (HEK293). It contains AA Ala 27 - Arg 191 (Accession # P15692-4).

Predicted N-terminus: His

Molecular Characterization

Poly-his
Avi
VEGF165(Ala 27 - Arg 191)P15692-4

This protein carries a polyhistidine tag at the N-terminus, followed by an Avi tag (Avitag™).

The protein has a calculated MW of 22.4 kDa (monomer). As a result of glycosylation, the protein migrates as 30 kDa (monomer) under reducing (R) condition, and 48-60 kDa (homodimer) under non-reducing (NR) condition (SDS-PAGE).

Biotinylation

Biotinylation of this product is performed using Avitag™ technology. Briefly, the single lysine residue in the Avitag is enzymatically labeled with biotin.

Biotin:Protein Ratio

The biotin to protein ratio is 0.5-1 as determined by the HABA assay.

Endotoxin

Less than 1.0 EU per μg by the LAL method.

Purity

>95% as determined by SDS-PAGE.

Formulation

Lyophilized from 0.22 μm filtered solution in PBS, pH7.4 . Normally trehalose is added as protectant before lyophilization.

Contact us for customized product form or formulation.

Reconstitution

Please see Certificate of Analysis for specific instructions.

For best performance, we strongly recommend you to follow the reconstitution protocol provided in the CoA.

Storage

For long term storage, the product should be stored at lyophilized state at -20°C or lower.

Please avoid repeated freeze-thaw cycles.

No activity loss is observed after storage at:

  1. 4-8°C for 12 months in lyophilized state;
  2. -70°C for 3 months under sterile conditions after reconstitution.
 

SDS-PAGE

Biotinylated Human VEGF165, His Tag (Cat. No. VE5-H82Q0) SDS-PAGE gel

Biotinylated Human VEGF165, His Tag on SDS-PAGE under reducing (R) and no-reducing (NR) conditions. The gel was stained overnight with Coomassie Blue. The purity of the protein is greater than 95%.

 

Bioactivity

Biotinylated Human VEGF165, His Tag (Cat. No. VE5-H82Q0) ELISA bioactivity

Immobilized Human VEGFR1/R2, Fc tag at 2 μg/mL (100 μL/well) can bind Biotinylated Human VEGF165 (Cat. No. VE5-H82Q0) with a linear range of 0.5-7.8 ng/mL (QC tested).

Request for protocol

Biotinylated Human VEGF165, His Tag (Cat. No. VE5-H82Q0) ELISA bioactivity

Immobilized Anti-VEGF MAb,Human IgG1 (Avastin) at 5 μg/mL (100 μL/well) can bind Biotinylated Human VEGF165 (Cat. No. VE5-H82Q0) with a linear range of 1-15.6 ng/mL (Routinely tested).

Request for protocol

Biotinylated Human VEGF165, His Tag (Cat. No. VE5-H82Q0) ELISA bioactivity

Immobilized biotinylated human VEGF165(Cat. No. VE5-H82Q0) on Streptavidin Chip , can bind Avastin with an affinity constant of 0.417nM as determined in SPR assay (Biacore T200) ( Routinely tested).

Request for protocol

 

Background

VEGF165 is the most abundant splice variant of VEGF-A. VEGF165 is produced by a number of cells including endothelial cells, macrophages and T cells. VEGF165 is involved in angiogenesis, vascular endothelial cell survival, growth, migration and vascular permeability. VEGF gene expression is induced by hypoxia, inflammatory cytokines and oncogenes. VEGF165 binds to heparan sulfate and is retained on the cell surface and in the extracellular matrix. VEGF165 binds to the receptor tyrosine kinases, VEGFR1 and VEGFR2. VEGF165 is the only splice variant that binds to co-receptors NRP-1 and NRP-2 that function to enhance VEGFR2 signaling. Binding of VEGF165 to VEGFR1 and VEGFR2 leads to activation of the PI3K/AKT, p38 MAPK, FAK and paxillin. VEGF plays a key role in tumor angiogenesis in many cancers.

References

Please contact us via TechSupport@acrobiosystems.com if you have any question on this product.


Limited Use License

The Biotin AviTag technology is covered by U.S. Pat. No: 5,874,239 and includes any and all materials, methods, kits and related derivatives claimed by this patent. The purchase of the Acrobiosystems’s Avitag™ proteins confers to the purchaser the limited right to use the Avitag™ technology for non-commercial, or research use, or for purposes of evaluating the Avitag™ technology.

Commercial use of the Avitag™ technology to manufacture a commercial product, or use of the Avitag™ technology to facilitate or advance research which will be applied to the development of a commercial product requires a license from Avidity, LLC. Examples of Commercial use include, but are not limited to biosensors, diagnostics, therapeutic applications, proximity assays, and drug screening assays.

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