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Your Position: 홈 > Protein > VEGF165 > VE5-H82Q0

Biotinylated Human VEGF165 Protein, His,Avitag™

INTRODUCTION
QC & Assay Protocol
Citation & Background
  • Synonym
    RP1-261G23.1,MGC70609,MVCD1,VEGFA,VPF
  • Source
    Biotinylated Human VEGF165, His,Avitag(VE5-H82Q0) is expressed from human 293 cells (HEK293). It contains AA Ala 27 - Arg 191 (Accession # P15692-4).
    Predicted N-terminus: His
    Request for sequence
  • Molecular Characterization
    VEGF165 Structure

    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). The protein migrates as 25-33 kDa (monomer) under reducing (R) condition, and 42-55 kDa (homodimer) under non-reducing (NR) condition (SDS-PAGE) due to glycosylation.

  • Labeling
    Biotinylation of this product is performed using Avitag™ technology. Briefly, the single lysine residue in the Avitag is enzymatically labeled with biotin.
  • Protein Ratio
    Passed as determined by the HABA assay / binding ELISA.
  • 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 with trehalose as protectant.

    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.

    This product is stable after storage at:

    1. -20°C to -70°C for 12 months in lyophilized state;
    2. -70°C for 12 months under sterile conditions after reconstitution.
SDS-PAGE
VEGF165 SDS-PAGE

Biotinylated Human VEGF165, His,Avitag on SDS-PAGE under reducing (R) and non-reducing (NR) conditions. The gel was stained with Coomassie Blue. The purity of the protein is greater than 95%.

Bioactivity-ELISA
VEGF165 ELISA

Immobilized Human VEGF R2, His Tag (Cat. No. KDR-H5227) at 2 μg/mL (100 μL/well) can bind Biotinylated Human VEGF165, His,Avitag (Cat. No. VE5-H82Q0) with a linear range of 2-39 ng/mL (QC tested).

Protocol
VEGF165 ELISA

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

Protocol
VEGF165 ELISA

Immobilized Human VEGF R2, His Tag (Cat. No. KDR-H5227) at 2 μg/mL (100 μL/well), can bind pre-mixed increasing concentrations of Bevacizumab and 0.05 μg/mL (100 μL/well) Biotinylated Human VEGF165, His,Avitag (Cat. No. VE5-H82Q0) with a half maximal inhibitory concentration (IC50) of 0.74 μg/mL (Routinely tested).

Protocol
Bioactivity-SPR
VEGF165 SPR

Immobilized Biotinylated Human VEGF165, His,Avitag (Cat. No. VE5-H82Q0) on CM5 Chip via Streptavidin, can bind Avastin with an affinity constant of 0.417 nM as determined in SPR assay (Biacore T200) (Routinely tested).

Protocol
Bioactivity-BLI
VEGF165 BLI

Loaded Biotinylated Human VEGF165, His,Avitag (Cat. No. VE5-H82Q0) on SA Biosensor, can bind Human Neuropilin-1, His Tag (Cat. No. NR1-H5228) with an affinity constant of 21.1 nM as determined in BLI assay (ForteBio Octet Red96e) (Routinely tested).

Protocol
VEGF165 BLI

Loaded Biotinylated Human VEGF165, His,Avitag (Cat. No. VE5-H82Q0) on SA Biosensor, can bind Human VEGF R1, His Tag with an affinity constant of 3.36 nM as determined in BLI assay (ForteBio Octet Red96e) (Routinely tested).

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.
  • Clinical and Translational Updates

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