Recombinant Anti-VEGFA antibody [EP1176Y] - BSA and Azide free (ab185238)

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)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52917).

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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52917).

Clone EP1176Y (ab185238) has been successfully conjugated by Abcam. This image was generated using Anti-VEGFA antibody [EP1176Y] (PE). Please refer to ab209439 for protocol details. 

Overlay histogram showing HeLa cells stained with ab209439 (red line). The cells were fixed with 4% formaldehyde (10 min) and then permeabilized with 90% methanol at -20°C for 15 min. The cells were then incubated in 1x PBS / 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (ab209439, 1/2500 dilution) for 30 min at 22°C.Isotype control antibody (black line) was rabbit IgG (monoclonal) Phycoerythrin (ab209478) used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20 mW Solid State Blue Laser (488nm) and 585/40 bandpass filter. 

Clone EP1176Y (ab185238) has been successfully conjugated by Abcam. This image was generated using Anti-VEGFA antibody [EP1176Y] (Alexa Fluor® 647). Please refer to ab206887 for protocol details.

ab206887 staining VEGF in HeLa cells. The cells were fixed with 4% formaldehyde (10 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab206887 at 1/100 dilution (shown in red) and ab195887, Mouse monoclonal to alpha Tubulin (Alexa Fluor^® 488), at 1/250 dilution (shown in green). Nuclear DNA was labelled with DAPI (shown in blue).

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

Clone EP1176Y (ab185238) has been successfully conjugated by Abcam. This image was generated using Anti-VEGFA antibody [EP1176Y] (Alexa Fluor® 488). Please refer to ab206886 for protocol details.

ab206886 staining VEGF in HeLa (Human epithelial cell line from cervix adenocarcinoma) cells. The cells were fixed with 4% formaldehyde (10 minutes), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1 hour. The cells were then incubated overnight at +4°C with ab206886 at 1/1000 dilution (shown in green) and ab195889, Mouse monoclonal to alpha Tubulin (Alexa Fluor^® 594), at 1/250 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

Intracellular 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). 

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52917). 

Immunohistochemistry (Paraffin-embedded sections) analysis of Human kidney tissue labeling VEGF with ab185238 at 1/250. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9.0. Anti-rabbit IgG H&L (HRP) (ab97051) was used as the secondary antibody. Counterstained with Hematoxylin.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52917).

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 22°C. The secondary antibody used was Alexa Fluor^® 488 goat anti-rabbit IgG (H&L) (ab150081) at 1/2000 dilution for 30 min at 22°C. 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 data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52917). 

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 data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52917).

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