The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/100 - 1/1000.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
Use at an assay dependent concentration.
1/250 - 1/500.
1/1000. Detects a band of approximately 23 kDa (predicted molecular weight: 27 kDa).
Growth factor active in angiogenesis, vasculogenesis and endothelial cell growth. Induces endothelial cell proliferation, promotes cell migration, inhibits apoptosis and induces permeabilization of blood vessels. Binds to the FLT1/VEGFR1 and KDR/VEGFR2 receptors, heparan sulfate and heparin. NRP1/Neuropilin-1 binds isoforms VEGF-165 and VEGF-145. Isoform VEGF165B binds to KDR but does not activate downstream signaling pathways, does not activate angiogenesis and inhibits tumor growth.
Isoform VEGF189, isoform VEGF165 and isoform VEGF121 are widely expressed. Isoform VEGF206 and isoform VEGF145 are not widely expressed.
Involvement in disease
Defects in VEGFA are a cause of susceptibility to microvascular complications of diabetes type 1 (MVCD1) [MIM:603933]. These are pathological conditions that develop in numerous tissues and organs as a consequence of diabetes mellitus. They include diabetic retinopathy, diabetic nephropathy leading to end-stage renal disease, and diabetic neuropathy. Diabetic retinopathy remains the major cause of new-onset blindness among diabetic adults. It is characterized by vascular permeability and increased tissue ischemia and angiogenesis.
Belongs to the PDGF/VEGF growth factor family.
Secreted. VEGF121 is acidic and freely secreted. VEGF165 is more basic, has heparin-binding properties and, although a signicant proportion remains cell-associated, most is freely secreted. VEGF189 is very basic, it is cell-associated after secretion and is bound avidly by heparin and the extracellular matrix, although it may be released as a soluble form by heparin, heparinase or plasmin.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-VEGFA antibody [EP1176Y] (ab52917)This image is courtesy of an anonymous Abreview
ab52917 staining VEGF in Mouse kidney tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde, permeabilized with 0.3% Triton X-100 and blocked with 5% serum for 45 minutes at 25°C. Samples were incubated with primary antibody (1/400 in 4% BSA + 5% serum in PBST) for 14 hours at 4°C. An Alexa Fluor® 546-conjugated Donkey anti-rabbit IgG polyclonal (1/300) was used as the secondary antibody.
ab52917 staining VEGF in NIH3T3 cells. The cells were fixed with 4% formaldehyde (10min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated with ab52917 at 5μg/ml and ab195884, at 1/250 dilution, overnight at +4°C, followed by a further incubation at room temperature for 1h with an Goat anti-Rabbit Alexa Fluor 488 secondary (ab150081) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.
Western blot - Anti-VEGFA antibody [EP1176Y] (ab52917)
All lanes : Anti-VEGFA antibody [EP1176Y] (ab52917) at 1 µg/ml
Lane 1 :Recombinant Human VEGFA protein (ab204773) Lane 2 :Recombinant mouse VEGFA protein (Active) (ab185265)
Lysates/proteins at 0.1 µg per lane.
Secondary All lanes : Rabbit IgG secondary antibody at 1/10000 dilution
This blot was produced using a 10% Bis-tris gel under the MES buffer system. The gel was run at 200V for 35 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% Milk before being incubated with ab52917 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution.
Overlay histogram showing NIH3T3 cells stained with ab52917 (red line). The cells were fixed with 4% formaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab52917, 1/1000 dilution) for 30 min at 22ºC. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150081) at 1/2000 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (ab172730, 0.1μg/1x106 cells used under the same conditions. Unlabelled sample (blue line) was also used as a control.
Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.
This antibody gave a positive signal in NIH3T3 cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used at 1/100 dilution.
This product has been referenced in:
Alshaker H et al. Combination of RAD001 (everolimus) and docetaxel reduces prostate and breast cancer cell VEGF production and tumour vascularisation independently of sphingosine-kinase-1. Sci Rep7:3493 (2017).
Read more (PubMed: 28615679) »
Wang H et al. Growth of MCF-7 breast cancer cells and efficacy of anti-angiogenic agents in a hydroxyethyl chitosan/glycidyl methacrylate hydrogel. Cancer Cell Int17:55 (2017).
Read more (PubMed: 28515673) »