All lanes : Anti-PP2A alpha + beta antibody [YE351] (ab32065) at 1/50000 dilution

Lane 1 : A431 whole cell lysate
Lane 2 : A549 whole cell lysate
Lane 3 : Caco-2 whole cell lysate
Lane 4 : T47D whole cell lysate
Lane 5 : C2C12 whole cell lysate
Lane 6 : HeLa whole cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

Predicted band size: 36 kDa
Observed band size: 36 kDa


Exposure time: 15 seconds

Blocking and dilution buffer: 5% NFDM/TBST.

All lanes : Anti-PP2A alpha + beta antibody [YE351] (ab32065) at 1/50000 dilution

Lane 1 : Jurkat whole cell lysate
Lane 2 : U-937 whole cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

Predicted band size: 36 kDa
Observed band size: 36 kDa


Exposure time: 5 seconds

Blocking and dilution buffer: 5% NFDM/TBST.

All lanes : Anti-PP2A alpha + beta antibody [YE351] (ab32065) at 1/100000 dilution

Lane 1 : Human fetal heart tissue lysate
Lane 2 : Human fetal kidney tissue lysate
Lane 3 : Human fetal spleen tissue lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : HRP-conjugated anti-rabbit IgG, specific to the non-reduced form of IgG at 1/1000 dilution

Predicted band size: 36 kDa
Observed band size: 36 kDa


Exposure time: 30 seconds

Blocking and dilution buffer: 5% NFDM/TBST.

Anti-PP2A alpha + beta antibody [YE351] (ab32065) at 1/100000 dilution + Human fetal brain tissue lysate at 10 µg

Secondary
HRP-conjugate anti-rabbit IgG, specific to the non-reduced form of IgG at 1/1000 dilution

Predicted band size: 36 kDa
Observed band size: 36 kDa


Exposure time: 15 seconds

Blocking and dilution buffer: 5% NFDM /TBST.

All lanes : Anti-PP2A alpha + beta antibody [YE351] (ab32065) at 1/100000 dilution

Lane 1 : Mouse brain tissue lysate
Lane 2 : Mouse heart tissue lysate
Lane 3 : Mouse kidney tissue lysate
Lane 4 : Mouse spleen tissue lysate
Lane 5 : Rat brain tissue lysate
Lane 6 : Rat heart tissue lysate
Lane 7 : Rat kidney tissue lysate
Lane 8 : Rat spleen tissue lysate
Lane 9 : C6 whole cell lysate
Lane 10 : Raw264.7 whole cell lysate
Lane 11 : PC-12 whole cell lysate
Lane 12 : NIH/3T3 whole cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

Predicted band size: 36 kDa
Observed band size: 36 kDa


Exposure time: 1 second

Blocking and dilution buffer: 5% NFDM /TBST.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human tonsil tissue labeling PP2A alpha + beta with ab32065 at a dilution of 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

Cytoplasm and weak nucleus staining is visible in lymphocytes of human tonsil.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse kidney tissue labelling PP2A alpha + beta with ab32065 at a dilution of 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

Cytoplasm staining is visible in epithelial cells of mouse kidney

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat kidney tissue labelling PP2A alpha + beta with ab32065 at a dilution of 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

Cytoplasm and weak nucleus staining is visible in epithelial cells of rat kidney.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse heart tissue labelling PP2A alpha + beta with ab32065 at a dilution of 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

Cytoplasm staining is visible in mouse heart.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat heart tissue labelling PP2A alpha + beta with ab32065 at a dilution of 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

Cytoplasm and nucleus staining is visible on rat heart.

ab32065 at 1/80 immunoprecipitating PP2A alpha + beta in HeLa whole cell lysate.

Lane 1 (input): HeLa whole cell lysate (10µg)

Lane 2 (+): ab32065 + HeLa whole cell lysate.

Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab32065 in HeLa (human cervix adenocarcinoma) whole cell lysate.

For western blotting, ab32065 (1/500) ab131366 VeriBlot for IP Detection Reagent (HRP) was used for detection (1/10000).

Blocking buffer and concentration: 5% NFDM/TBST.

Diluting buffer and concentration: 5% NFDM /TBST.

Observed band: 37kDa.

 

Dot blot analysis of PP2A-alpha peptide (Lane 1) and PP2A-beta peptide (Lane 2) labelling PP2A alpha+beta with ab32065 at a dilution of 1/1000. A HRP-conjugated anti-rabbit IgG, specific to the non-reduced form of IgG was used as the secondary antibody at a dilution of 1/1000.

Blocking and dilution buffer: 5% NFDM/TBST.

Exposure time: 3 minutes.