VEGF-A
A microtiter plate was coated overnight with recombinant human VEGF-A (PHP293) at a concentration of 5 µg/ml. After washing and blocking with PBST+5% BSA, 10% human serum was added spiked with increasing concentrations of ranibizumab. Detection was performed using Human Anti-Ranibizumab Antibody, clone AbD29928 (HCA304) at a concentration of 2 µg/ml and a rat anti-DYKDDDDK-tag antibody in HISPEC Assay Diluent (BUF049A) followed by QuantaBlu Fluorogenic Peroxidase Substrate. Data are shown as the mean of three measurements.
A microtiter plate was coated overnight with recombinant human VEGF-A (PHP293) at a concentration of 5 µg/ml. After washing and blocking with PBST+5% BSA, 10% human serum was added spiked with increasing concentrations of bevacizumab. Detection was performed using Human Anti-Ranibizumab Antibody, clone AbD29865 (HCA307, cross-reactive with bevacizumab) at a concentration of 2 μg/ml and a rat anti-DYKDDDDK-tag antibody in HISPEC Assay Diluent (BUF049A) followed by QuantaBlu Fluorogenic Peroxidase Substrate. Data are shown as the mean of three measurements.
Recombinant Human VEGF-A
Vascular endothelial growth factor alpha forms a homodimer and is a potent stimulator of angiogenesis of both normal and cancerous cells. It acts as a regulator of vasculogenesis, angiogenesis and endothelial cell growth. VEGF-A is secreted by many different cell types such as endothelial cells (Nissen et al. 1998), smooth muscle cells (Brogi et al. 1994), neutrophils (Gaudry et al. 1997), platelets (Banks et al. 1998), macrophages and 60% of all tumours (Berse et al. 1992).
The sequence of this recombinant protein product is amino acid (aa) 27 - aa 141 + CDKPRR but deficient from aa 142-226, and it is closest to isoform 121. Isoform 121 and 111 lack exons 6 and 7, and for this reason neither of these isoforms bind to the extracellular matrix (Krilleke et al. 2017). This product has been demonstrated for use, in a pharmacokinetic (PK) antigen capture ELISA with the monoclonal antibody drugs ranibizumab and bevacizumab using anti-ranibizumab antibodies for detection.
Vascular endothelial growth factor alpha forms a homodimer and is a potent stimulator of angiogenesis of both normal and cancerous cells. It acts as a regulator of vasculogenesis, angiogenesis and endothelial cell growth. VEGF-A is secreted by many different cell types such as endothelial cells (Nissen et al. 1998), smooth muscle cells (Brogi et al. 1994), neutrophils (Gaudry et al. 1997), platelets (Banks et al. 1998), macrophages and 60% of all tumours (
The sequence of this recombinant protein product is amino acid (aa) 27 - aa 141 + CDKPRR but deficient from aa 142-226, and it is closest to isoform 121. Isoform 121 and 111 lack exons 6 and 7, and for this reason neither of these isoforms bind to the extracellular matrix (Krilleke et al. 2017). This product has been demonstrated for use, in a pharmacokinetic (PK) antigen capture ELISA with the monoclonal antibody drugs ranibizumab and bevacizumab using anti-ranibizumab antibodies for detection.
Product Details
- Target Species
- Human
- Product Form
- Purified recombinant protein - lyophilized
- Reconstitution
- Centrifuge vial prior to reconstitution. Reconstitute to 0.5 mg/ml by adding 0.4 ml ddH2O.
Care should be taken during reconstitution as the protein may appear as a film at the bottom of the vial. Mix gently after reconstitution. Do not vortex. - Preparation
- Recombinant protein expressed in E. coli and purified by affinity chromatography
- Buffer Solution
- 20 mM Phosphate Buffer, 0.1 M Sodium Chloride
- Preservative Stabilisers
- 1% Trehalose
- Purity
- ≥90% determined by SDS-PAGE under reducing conditions and visualized by coomassie blue staining
- Approx. Protein Concentrations
- 0.5 mg/ml after reconstitution
- Protein Molecular Weight
- Predicted 14 kDa. The apparent molecular mass of the human VEGF-A monomer is approximately 17 kDa determined by SDS-PAGE under reducing conditions, and 17 kDa (monomer) and 30 kDa (dimer) under non-reducing conditions.
Storage Information
- Storage
- Prior to reconstitution store at -70oC. Following reconstitution store at -20oC.
This product should be stored undiluted.
Storage in frost-free freezers is not recommended. Avoid repeated freezing and thawing as this may denature the protein. - Guarantee
- Guaranteed for 3 months from the date of reconstitution or until the date of expiry, whichever comes first. Please see label for expiry date.
More Information
- UniProt
- P15692
- Regulatory
- For research purposes only
Applications of VEGF-A
| Application Name | Verified | Min Dilution | Max Dilution |
|---|---|---|---|
| ELISA |
Where this product has not been tested for use in a particular technique this does not necessarily exclude its use in such procedures. Suggested working dilutions are given as a guide only. It is recommended that the user titrates the product for use in their own system using appropriate negative/positive controls.
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Useful Reagents Available
| Description | Product Code | Applications | Pack Size | List Price | Quantity |
|---|---|---|---|---|---|
| Human anti Ranibizumab (Drug/Target Complex) | HCA304 | E | 0.1 mg |
|
|
| Human anti Ranibizumab | HCA307 | E | 0.1 mg |
|
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