ab52619 staining PPP1CA + 1CB in the HepG2 cell line by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 4% Paraformaldehyde permeabilized with 0.1% Triton X-100. Samples were incubated with primary antibody (1/50). ab150077(1/500) an Alexa Fluor^®488-conjugated Goat anti-rabbit IgG was used as the secondary antibody. Nuclei were counterstained with DAPI.

All lanes : Anti-PPP1CA + PPP1CB antibody [EP1511Y] (ab52619) at 1/100000 dilution

Lane 1 : Mouse Brain lysate
Lane 2 : Rat Brain lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), HRP-conjugated at 1/1000 dilution

Predicted band size: 37 kDa

ab52619 staining PP1CA + 1CB in Human cerebrum cortex tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed and paraffin-embedded, antigen retrieval was by heat mediation in Tris/EDTA buffer pH9. Samples were incubated with primary antibody (1/50). An undiluted HRP-conjugated mouse anti-rabbit IgG was used as the secondary antibody. Tissue counterstained with Hematoxylin. PBS was used in the negative control rather than the Primary antibody.

Overlay histogram showing HeLa cells stained with ab52619 (red line) at 1/150 dilution. The cells were fixed with 80% methanol. The secondary antibody used was a FITC conjugated goat anti-rabbit IgG at 1/150 dilution. Isotype control antibody (black line) was rabbit monoclonal IgG used under the same conditions. Cells also incubated without primary antibody and secondary antibody (blue line)

ab52619 (purified) at 1/30 immunoprecipitating PPP1CA + 1CB in Jurkat cell lysate. For western blotting, a HRP-conjugated Goat anti-rabbit IgG, specific to the non-reduced form of IgG was used as the secondary antibody (1/1000).

Blocking buffer and concentration: 5% NFDM/TBST.

Diluting buffer and concentration: 5% NFDM /TBST.

All lanes : Anti-PPP1CA + PPP1CB antibody [EP1511Y] (ab52619) at 1/100000 dilution

Lane 1 : T47-D cell Lysate
Lane 2 : HeLa cell Lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), HRP-conjugated at 1/1000 dilution

Predicted band size: 37 kDa

Overlay histogram showing HeLa cells stained with ab52619, unpurified (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 (ab52619, 1/100 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight^® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed.

All lanes : Anti-PPP1CA + PPP1CB antibody [EP1511Y] (ab52619) at 1/20000 dilution

Lane 1 : SKBR-3 cell lysate
Lane 2 : Jurkat cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), HRP-conjugated at 1/1000 dilution

Predicted band size: 37 kDa