Anti-VEGFA antibody [EP1176Y] - BSA and Azide free

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-VEGFA antibody [EP1176Y] - BSA and Azide free (AB185238)

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).

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-VEGFA antibody [EP1176Y] - BSA and Azide free (AB185238)

This data was developed using ab52917, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of formalin fixed paraffin embedded human kidney labelling VEGFA with ab52917 at a concentration of 2µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with a ChromoMap DAB IHC Detection Kit followed by anti rabbit HQ and anti HQ HRP detection. Heat mediated antigen retrieval was performed with DISCOVERY cell conditioning solution (CC1) 100°C, pH8.5 for 64mins.

ab52917 Anti-VEGFA antibody [EP1176Y] was incubated for 16mins at 37°C. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.

Customers are encouraged to optimise antigen retrieval conditions, antibody concentration, incubation times and temperature for best results in their own IHC assay workflow (automated and manual).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - 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).

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-VEGFA antibody [EP1176Y] - BSA and Azide free (AB185238)

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).

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-VEGFA antibody [EP1176Y] - BSA and Azide free (AB185238)

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.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-VEGFA antibody [EP1176Y] - BSA and Azide free (AB185238)

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).

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-VEGFA antibody [EP1176Y] - BSA and Azide free (AB185238)

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).

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-VEGFA antibody [EP1176Y] - BSA and Azide free (AB185238)

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).

• IHC-P

AbReview38570****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-VEGFA antibody [EP1176Y] - BSA and Azide free (AB185238)

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).

This image is courtesy of an anonymous Abreview.