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Synthetic peptide corresponding to residues in N terminus of human VEGF Receptor 1
This product is a recombinant rabbit monoclonal antibody.
Produced using Abcam’s RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5,675,063 and/or 7,429,487.
Alternative versions available:
Our Abpromise guarantee covers the use of ab32152 in the following tested applications.
|WB||1/1000 - 1/5000. Predicted molecular weight: 151 kDa.|
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
Immunohistochemistry (PFA perfusion fixed frozen sections) analysis of mouse brain tissue section (15 days old wild-type mouse embryonic brain, 16 micron) labeling VEGF Receptor 1 with ab32152 at 1/300 dilution. Tissue was fixed with formaldehyde and permeabilized with Triton X100. Heat mediated antigen retrieval was performed using 10mM citrate buffer, pH 6. A polyclonal donkey anti-Rabbit IgG (H+L) (Alexa Fluor® 488) secondary antibody was used at 1/500 dilution.
Immunocytochemistry/ Immunofluorescence analysis of mouse cardiac cells labeling VEGF Receptor 1 with ab32152. An Alexa Fluor-conjugated secondary antibody was used. Blue: DAPI counterstaining, Green : GFP, Red: VEGF Receptor 1.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human abdominal aortic aneurysm (AAA) wall tissue sections labeling VEGF Receptor 1 with ab32152 at 1/100 dilution.
Resected aortic tissues were immersed in 10% neutral buffered formalin for at least 24 h for immunohistochemical staining. Tissue sample was embedded in paraffin; 4 µm sections were cut and mounted onto MAS-coated slides. The sections were deparaffinized, dehydrated, and boiled in a pressure cooker in 0.01 M citric acid buffer (pH 6.0) for 20 min. The sections were washed with phosphate-buffered saline and incubated with 3% H2O2 in absolute methanol for 5 min to inhibit any endogenous peroxidase activity. Sections were preincubated with 3% normal goat serum for 20 min to minimize nonspecific binding to VEGF Receptor 1, and incubated with ab32152 at 4°C overnight in a moist chamber. The section was washed with phosphate-buffered saline and then incubated with the appropriate secondary antibody for 30 min at room temperature. Staining was visualized with Vector DAB, and tissue section was then counterstained with hematoxylin.
Overlay histogram showing A431 cells stained with ab32152 (red line). The cells were fixed with 80% methanol (5 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 (ab32152, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used a goat anti-rabbit DyLight® 488 goat (IgG, H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (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.
Flow Cytometry analysis of A7r5 (left) and A10 (middle) and MVSC (right) cells labeling VEGF Receptor 1 with ab32152. Open curves represent negative control samples; red filled curves represent samples stained with ab32152.
Cells were cultured until confluency in T75 flasks and then trypsinised in 2x trypsin/0.53 mM EDTA at 37°C. Cells were resuspended in media, counted, and then fixed for 20 mins at 4°C. Cells were then washed by centrifugation at 500 g for 3 mins. Following washing, cells were resuspended in solution containing 1 μg of Anti-VEGF Receptor 1 antibody [Y103] (ab32152) and incubated at 4°C for 30 mins. Following washing, cells were resuspended in solution containing 1 μg of appropriate enzyme-linked secondary antibody and incubated at 4°C for 30 mins. Cells were then washed in solution and resuspended in a final volume of 500 μL of solution.