Recombinant Anti-VAMP2 antibody [EPR12790] - BSA and Azide free (ab214590)

ab181869 staining VAMP2 in U87-MG (human glioblastoma) cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 100% methanol. Samples were incubated with primary antibody at a dilution of 1/500. A goat anti rabbit IgG (Alexa Fluor® 488) (ab150077) was used as the secondary antibody at a dilution of 1/1000. DAPI was used as a nuclear counterstain.

Negative control 1:PBS only.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab181869).

ab181869 (purified) at 1/150 immunoprecipitating VAMP2 in Human cerebellum whole cell lysate. 10 ug of cell lysate was present in the input. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution. A rabbit monoclonal IgG (ab172730) was used intead of ab128913 as a negative control (Lane 3).

Blocking buffer and concentration: 5% NFDM/TBST.

Diluting buffer and concentration: 5% NFDM /TBST.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab181869).

Clone EPR12790 (ab214590) has been successfully conjugated by Abcam. This image was generated using Anti-VAMP2 antibody [EPR12790] (PE). Please refer to ab214529 for protocol details.

Overlay histogram showing U-87MG cells stained with ab214529 (red line). The cells were fixed with 4% formaldehyde (10 min) and then permeabilized with 0.1% PBS-Triton X-100 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 (ab214529, 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 50mW Yellow/Green laser (561nm) and 586/15 bandpass filter.

This antibody gave a positive signal in U-87MG cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Triton X-100 for 15 min used under the same conditions.

Clone EPR12790 (ab214590) has been successfully conjugated by Abcam. This image was generated using Anti-VAMP2 antibody [EPR12790] (Alexa Fluor® 647). Please refer to ab198949 for protocol details.

ab198949 staining VAMP2 in U87MG cells. The cells were fixed with 100% methanol (5 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 ab198949 at a 1/100 dilution (shown in red) and ab195887, Mouse monoclonal to alpha Tubulin (Alexa Fluor^® 488), at a 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 Cytometry analysis of SH-SY5Y cells labelling VAMP2 with purified ab181869 at 1/100 (red). Cells were fixed with 4% paraformaldehyde. A FITC-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab181869).

Immunocytochemistry/Immunofluorescence analysis of U87-MG (human glioblastoma) cells labelling VAMP2 with purified ab181869 at 1/250. Cells were fixed with 100% methanol and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor^® 594-conjugated goat anti-mouse IgG (1/1000) were also used.

Control 1: primary antibody (1/100) and secondary antibody, ab150120, an Alexa Fluor^® 594-conjugated goat anti-mouse IgG (1/500).

Control 2: ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/500).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab181869).

Western blot analysis of immunoprecipitation pellet from Human cerebellum lysate immunoprecipitated using ab181869 at 1/70 dilution.
Secondary: Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugate at 1/1000 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab181869).

Flow cytometry analysis of Jurkat cells fixed with 2% paraformaldehyde labeling VAMP2 with unpurified ab181869 at 1/150 dilution followed by Goat anti rabbit IgG (FITC) at 1/150 dilution. Rabbit monoclonal IgG was used as an isotype control.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab181869).

Immunofluorescent analysis of SH-SYSY cells (4% paraformaldehyde-fixed) labeling VAMP2 with unpurified ab181869 at 1/500 dilution followed by Goat anti rabbit IgG (Alexa Fluor^®488) at 1/200 dilution and counterstained with DAPI.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab181869).

Immunofluorescent analysis of U87-MG cells (4% paraformaldehyde-fixed)  labeling VAMP2 with ab181869 at 1/500 dilution followed by Goat anti rabbit IgG (Alexa Fluor®488) at 1/200 dilution and counterstained with Dapi.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab181869).