All lanes : Anti-PPP1CB antibody [EP1804Y] (ab53315) at 1/50000 dilution (purified)

Lane 1 : Jurkat (Human T cell leukemia T lymphocyte) whole cell lysates
Lane 2 : Mouse brain lysates
Lane 3 : Rat brain lysates

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Predicted band size: 37 kDa
Observed band size: 38 kDa why is the actual band size different from the predicted?

Blocking and diluting buffer: 5% NFDM/TBST

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human cervical carcinoma tissue sections labeling Protein phosphatase 1 beta with purified ab53315 at 1:400 dilution (1 μg/ml). Heat mediated antigen retrieval was performed using EDTA Buffer, pH 9.0. Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control.

Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling Protein phosphatase 1 beta with purified ab53315 at 1:40 dilution (10 ug/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 - 90% methanol. Unlabeled control - Rabbit monoclonal IgG (Black). Cell without incubation with primary antibody and secondary antibody (Blue).

ab53315 (purified) at 1:100 dilution (2ug) immunoprecipitating Protein phosphatase 1 beta in Human fetal brain lysate.

Lane 1 (input): Human fetal brain lysate 10ug

Lane 2 (+): ab53315 & Human fetal brain lysate

Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab53315 in Human fetal brain lysate

For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1:1000 dilution.

Blocking and diluting buffer: 5% NFDM/TBST.

Unpurified ab53315 staining Protein Phosphatase 1 beta antibody in HeLa (human cervix adenocarcinoma) cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 4% Paraformaldehyde and permeabilized with 0.1% Triton X-100. 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 and the negative control was PBS only.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Rat kidney tissue sections labeling Protein phosphatase 1 beta with purified ab53315 at 1:400 dilution (1 μg/ml). Heat mediated antigen retrieval was performed using EDTA Buffer, pH 9.0. Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Mouse kidney tissue sections labeling Protein phosphatase 1 beta with purified ab53315 at 1:400 dilution (1 μg/ml). Heat mediated antigen retrieval was performed using EDTA Buffer, pH 9.0. Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control.

Anti-PPP1CB antibody [EP1804Y] (ab53315) at 1/20000 dilution (unpurified) + Jurkat cells at 20 µg

Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/1000 dilution

Predicted band size: 37 kDa

Immunocytochemistry/ Immunofluorescence analysis of MCF-7 (Human breast adenocarcinoma epithelial cell) cells labeling Protein phosphatase 1 beta with Purified ab53315 at 1:250 dilution (1.6μg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 μg/ml). ab150077 Goat anti rabbit IgG(Alexa Fluor® 488) was used as the secondary antibody at 1:1000 dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

Immunohistochemical analysis of paraffin-embedded sections of human cervix carcinoma labelling Protein Phosphatase 1 beta with unpurified ab53315 at 1:250 dilution. Prediluted ImmunoHistoprobe (ready to use) HRP Polymer for Rabbit IgG was used as a secondary antibody.

Counter stain = Hematoxylin. Heat mediated antigen retrieval using EDTA buffer at pH 9 was performed.

Immunohistochemical analysis of paraffin-embedded sections of mouse cardiac muscle labelling Protein Phosphatase 1 beta with unpurified ab53315 at 1:250 dilution. Prediluted ImmunoHistoprobe (ready to use) HRP Polymer for Rabbit IgG was used as a secondary antibody.

Counter stain = Hematoxylin. Heat mediated antigen retrieval using EDTA buffer at pH 9 was performed.

Immunohistochemical analysis of paraffin-embedded sections of rat colon labelling Protein Phosphatase 1 beta with unpurified ab53315 at 1:250 dilution. Prediluted ImmunoHistoprobe (ready to use) HRP Polymer for Rabbit IgG was used as a secondary antibody.

Counter stain = Hematoxylin. Heat mediated antigen retrieval using EDTA buffer at pH 9 was performed.

Flow Cytometry analysis of HeLa cells labelling Protein Phosphatase 1 beta with unpurified ab53315 at 1:20 dilution. Goat anti rabbit IgG (H&L) (FITC) at a dilution of 1:150 used as the secondary antibody. Isotype control = Rabbit monoclonal IgG. Fixative=2% paraformaldehyde.