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ActiveMax® Recombinant Human VEGF165 (HPLC-verified)  pdf  pdf  pdf


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VE5-H4210-50ug
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Synonym

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

Source

ActiveMax® Human VEGF165 (VE5-H4210) is expressed from human 293 cells (HEK293). It contains AA Ala 27 - Arg 191 (Accession # NP_001165097).

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 (homodimer) under non-reducing (NR) condition on SDS-PAGE gel.

Endotoxin

Less than 1.0 EU per μg by the LAL method.

 

Purity

>98% as determined by SDS-PAGE.

>95% as determined by SEC-HPLC.

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

ActiveMax® Human VEGF165 (Catalog # VE5-H4210) SDS-PAGE gel

ActiveMax® 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 98%.

SEC-HPLC

ActiveMax® Human VEGF165 (Catalog # VE5-H4210) HPLC images

A SEC-HPLC analysis showing 95% of ActiveMax® Human VEGF165 (VE5-H4210) present as active homodimers.

 

Bioactivity

ActiveMax® Human VEGF165 (Catalog # VE5-H4210) ELISA bioactivity

(1) Immobilized ActiveMax® Human VEGF165 (Catalog # VE5-H4210, 1 ng/well) can bind Human VEGF R1, His Tag (Catalog # VE1-H5220) with a linear range of 4-30 ng/mL (Routinely tested).

Request for protocol

(3) Measured by its ability to bind Human VEGF R1, His Tag (Cat# VE1-H5220) in the SPR assay (Biacore 2000) with an estimated KD of 0.1nM (Routinely tested).

ActiveMax® Human VEGF165 (Catalog # VE5-H4210) ELISA bioactivity

(2) Immobilized ActiveMax® Human VEGF165 (Catalog # VE5-H4210) at 2 μg/mL can bind VEGFR2/R3-Fc with a linear range of 0.6-2.5 ng/mL (QC tested).

Request for protocol

(4) Measured by its ability to bind Human VEGF R2, His Tag (Cat# KDR-H5227) in the SPR assay (Biacore 2000) with an estimated KD < 50nM (Routinely tested).

 

Amino Acid Sequence

APMAEGGGQNHHEVVKFMDVYQRSYCHPIETLVDIFQEYPDEIEYIFKPSCVPLMRCGGCCNDEGLECVPTEESNITMQIMRIKPHQGQHIGEMSFLQHNKCECRPKKDRARQENPCGPCSERRKHLFVQDPQTCKCSCKNTDSRCKARQLELNERTCRCDKPRR

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.