MABSol® Biotinylated Human VEGF165, epitope tag free, primary amine labeling (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® Human VEGF165, Tag Free (HPLC-verified) (VE5-H4210).
Predicted N-terminus: Ala 27
This protein carries no "tag".
The protein has a calculated MW of 19.2 kDa. As a result of glycosylation, the protein migrates as 25-30 kDa (monomer) under reducing (R) condition, and 43-50 kDa under non-reducing (NR) condition (SDS-PAGE).
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.
The biotin to protein ratio is 3-5 as determined by the HABA assay.
Less than 1.0 EU per μg by the LAL method.
>95% as determined by reduced SDS-PAGE.
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.
Please see Certificate of Analysis for specific instructions.
For best performance, we strongly recommend you to follow the reconstitution protocol provided in the CoA.
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:
- -20°C to -70°C for 12 months in lyophilized state;
- -70°C for 3 months under sterile conditions after reconstitution.
Biotinylated Human VEGF165, epitope tag free, primary amine labeling on SDS-PAGE under reducing (R) and non-reducing (NR) conditions. The gel was stained overnight with Coomassie Blue. The purity of the protein is greater than 95%.
Immobilized human VEGFR1/R2, Fc Tag at 2 μg/mL (100 μL/well) can bind Biotinylated Human VEGF165, epitope tag free, primary amine labeling (Cat. No. VE5-H8210) with a linear range of 0.4-3 ng/mL (QC tested).
Immobilized Human Neuropilin-1, His Tag (Cat. No. NR1-H5228) at 2 μg/mL (100 μL/well) can bind Biotinylated Human VEGF165, epitope tag free, primary amine labeling (Cat. No. VE5-H8210) with a linear range of 5-40 ng/mL (Routinely tested).
Immobilized Anti-VEGF MAb, Human IgG1 (Avastin) at 5 μg/mL (100 μL/well) can bind Biotinylated Human VEGF165, epitope tag free, primary amine labeling (Cat. No. VE5-H8210) with a linear range of 0.2-2 ng/mL (Routinely tested).
Authors: Mcgonigle Sharon，Majumder Utpal，Postema Maarten H. D.
Journal: US20200206312A1 2020
Application: Binding Assay
Authors: McGonigle S, et al
Journal: Cell Commun Signal 43633
Authors: F Corti, et al
Journal: Nat Commun 43560
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.