Anti-cAMP Protein Kinase Catalytic subunit antibody [EP2102Y]

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-cAMP Protein Kinase Catalytic subunit antibody [EP2102Y] (AB76238)

Immunohistochemical (Formalin/PFA-fixed paraffin-embedded) analysis of ab76238 (unpurified), at a 1/100 dilution, staining human cAMP Protein Kinase Catalytic Subunit in testis.

Heat mediated antigen retrieval was performed before commencing with IHC staining protocol.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-cAMP Protein Kinase Catalytic subunit antibody [EP2102Y] (AB76238)

Intracellular Flow Cytometry analysis of MCF-7 (Human breast adenocarcinoma epithelial cell) cells labeling cAMP Protein Kinase Catalytic subunit with purified ab76238 at 1/80 dilution (10 μg/ml) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-cAMP Protein Kinase Catalytic subunit antibody [EP2102Y] (AB76238)

Overlay histogram showing HeLa cells stained with ab76238 (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 (ab76238, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1μg/1x10⁶ cells) used under the same conditions. Unlabelled sample (blue line). 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 HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-cAMP Protein Kinase Catalytic subunit antibody [EP2102Y] (AB76238)

Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling cAMP Protein Kinase Catalytic subunit with purified ab76238 at 1 : 100 dilution (8.0 µg/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). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1 : 1000 (2 µg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-cAMP Protein Kinase Catalytic subunit antibody [EP2102Y] (AB76238)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human thyroid carcinoma tissue sections labeling cAMP Protein Kinase Catalytic subunit with purified ab76238 at 1/750 dilution. Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

• IP

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Immunoprecipitation - Anti-cAMP Protein Kinase Catalytic subunit antibody [EP2102Y] (AB76238)

ab76238 (purified) at 1/40 dilution (2 µg) immunoprecipitating cAMP Protein Kinase Catalytic subunit in MCF-7 whole cell lysate.
Lane 1 (input) : MCF-7 (Human breast adenocarcinoma epithelial cell) whole cell lysate 10 µg
Lane 2 (+) : ab76238 & MCF-7 whole cell lysate
Lane 3 (-) : Rabbit monoclonal IgG (ab172730) instead of ab76238 in MCF-7 whole cell lysate
For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/1000 dilution.
Blocking and diluting buffer : 5% NFDM/TBST.

All lanes:

Immunoprecipitation - Anti-cAMP Protein Kinase Catalytic subunit antibody [EP2102Y] (ab76238)

Predicted band size: 41 kDa

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• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-cAMP Protein Kinase Catalytic subunit antibody [EP2102Y] (AB76238)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse cerebrum tissue sections labeling cAMP Protein Kinase Catalytic subunit with purified ab76238 at 1/750 dilution. Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-cAMP Protein Kinase Catalytic subunit antibody [EP2102Y] (AB76238)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat stomach tissue sections labeling cAMP Protein Kinase Catalytic subunit with purified ab76238 at 1/750 dilution. Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

• WB

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Western blot - Anti-cAMP Protein Kinase Catalytic subunit antibody [EP2102Y] (AB76238)

All lanes:

Western blot - Anti-cAMP Protein Kinase Catalytic subunit antibody [EP2102Y] (ab76238) at 1/200000 dilution

Lane 1:

HeLa cell lysate at 10 µg

Lane 2:

MCF-7 cell lysate at 10 µg

Secondary

All lanes:

goat anti-rabbit HRP at 1/1000 dilution

Predicted band size: 41 kDa

Observed band size: 42 kDa

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• WB

Lab

Western blot - Anti-cAMP Protein Kinase Catalytic subunit antibody [EP2102Y] (AB76238)

All lanes:

Western blot - Anti-cAMP Protein Kinase Catalytic subunit antibody [EP2102Y] (ab76238) at 1/20000 dilution

Lane 1:

MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysates at 15 µg

Lane 2:

NIH/3T3 (Mouse embryonic fibroblast) whole cell lysates at 15 µg

Lane 3:

C6 (Rat glial tumor glial cell) whole cell lysates at 15 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 41 kDa

Observed band size: 42 kDa

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