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Synthetic peptide within Human VEGFA aa 200-300 (C terminal). The exact sequence is proprietary.
A trial size is available to purchase for this antibody.
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
This product is a recombinant rabbit monoclonal antibody.
Our Abpromise guarantee covers the use of ab52917 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
The mouse recommendation is based on the WB results. We do not guarantee IHC-P for mouse.
|ICC/IF||1/250 - 1/500.|
|WB||1/10000. Detects a band of approximately 23 kDa (predicted molecular weight: 27 kDa).
This antibody fails to detect endogenous natural samples in WB.
Purified 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 purified 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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human kidney tissue sections labeling VEGFA with Purified ab52917 at 1:100 dilution (2.96 µg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0)
Flow Cytometry analysis of NIH/3T3 (Mouse embryonic fibroblast) cells labeling VEGFA with purified ab52917 at 1:30 dilution (10 µg/ml) (red). Cells were fixed with 4% Paraformaldehyde. A Goat anti rabbit IgG (Alexa Fluor® 488) secondary antibody was used at 1:2000 dilution. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).
Unpurified 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.
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 unpurified 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 (unpurified 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.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"