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Biotinylated Human VEGF165, epitope tag free, ultra sensitivity (primary amine labeling)  pdf  pdf  pdf


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Synonym

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

Source

MABSol® Biotinylated Human VEGF165 (VE5-H8210) is expressed from human HEK293 cells. It contains AA Ala 27 - Arg 191 (Accession # NP_001165097). It is the biotinylated form of ActiveMax® Recombinant Human VEGF165 (Cat. No. VE5-H4210).

Predicted N-terminus: Ala 27

Molecular Characterization

VEGF165(Ala 27 - Arg 191)NP_001165097

This protein carries no "tag".

The protein has a calculated MW of 19 kDa (monomer). As a result of glycosylation, the protein migrates as 24 kDa (monomer) under reducing (R) condition, and 43-50 kDa under non-reducing (NR) condition (SDS-PAGE).

Biotinylation

The primary amines in the side chains of lysine residues and the N-terminus of the protein are conjugated with biotins using standard chemical labeling method. A standard biotin reagent (13.5 angstroms) is used in this product.

Biotin:Protein Ratio

The biotin to protein ratio is 3-5 as determined by the HABA assay.

Endotoxin

Less than 1.0 EU per μg by the LAL method.

Purity

>95% as determined by reduced 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 (Cat. No. VE5-H8210) SDS-PAGE gel

Biotinylated Human VEGF165 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 (Cat. No. VE5-H8210) ELISA bioactivity

Immobilized human VEGFR1/R2, Fc tag at 2μg/mL (100 μL/well) can bind Biotinylated Human VEGF (Cat. No. VE5-H8210) with a linear range of 0.39-3.1 ng/mL (QC tested).

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Biotinylated Human VEGF165 (Cat. No. VE5-H8210) ELISA bioactivity

Immobilized Human Neuropilin-1, His Tag (Cat. No. NR1-H5228) at 2 μg/mL (100 μL/well) can bind Biotinylated Human VEGF165 (Cat. No. VE5-H8210) with a linear range of 5-40 ng/mL (Routinely tested).

Request for protocol

Biotinylated Human VEGF165 (Cat. No. VE5-H8210) ELISA bioactivity

Immobilized Anti-VEGF MAb,Human IgG1 (Avastin) at 5μg/mL (100 μL/well) can bind Biotinylated Human VEGF165 (Cat. No. VE5-H8210) with a linear range of 0.24-1.95 ng/mL (Routinely tested).

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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.

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