Anti-GSK3 alpha antibody [EP793Y]

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GSK3 alpha antibody [EP793Y] (AB40870)

ab4870 at 1/50 dilution, staining GSK3 alpha in Human breast carcinoma tissue by Immunohistochemistry, Paraffin embedded section.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-GSK3 alpha antibody [EP793Y] (AB40870)

Immunocytochemistry/ Immunofluorescence analysis of HeLa (human cervix adenocarcinoma epithelial cell) cells labeling GSK3 alpha with purified ab40870 at 1/200 dilution (10 μg/mL). Cells were fixed in 100% Methanol. 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.

• WB

Lab

Western blot - Anti-GSK3 alpha antibody [EP793Y] (AB40870)

Lanes 1-4 : Merged signal (red and green). Green - ab40870 observed at 51 kDa. Red - loading control ab8245 observed at 36 kDa.

ab40870 Anti-GSK3 alpha antibody [EP793Y] was shown to specifically react with GSK3 alpha in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab266742 (knockout cell lysate ab257979) was used. Wild-type and GSK3 alpha knockout samples were subjected to SDS-PAGE. ab40870 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-GSK3 alpha antibody [EP793Y] (ab40870) at 1/1000 dilution

Lane 1:

Wild-type HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 20 µg

Lane 2:

Western blot - Human GSK3A (GSK3 alpha) knockout HEK-293T cell lysate (<a href='/en-us/products/cell-lysates/human-gsk3a-gsk3-alpha-knockout-hek-293t-cell-lysate-ab257979'>ab257979</a>) at 20 µg

Lane 3:

HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg

Lane 4:

HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution

Predicted band size: 51 kDa

Observed band size: 51 kDa

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• WB

Supplier Data

Western blot - Anti-GSK3 alpha antibody [EP793Y] (AB40870)

Lane 1 : Wild-type HAP1 whole cell lysate (20 μg)
Lane 2 : GSK3 alpha knockout HAP1 whole cell lysate (20 μg)
Lane 3 : HeLa whole cell lysate (20 μg)
Lane 4 : HepG2 whole cell lysate (20 μg)

Lanes 1 - 4 : Merged signal (red and green). Green - ab40870 observed at 51 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab40870 was shown to specifically react with GSK3 alpha in wild-type HAP1 cells. No band was observed when GSK3 alpha knockout samples were examined. Wild-type and GSK3 alpha knockout samples were subjected to SDS-PAGE. ab40870 and ab8245 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at a 1/5000 dilution and 1/10,000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed ab216776 secondary antibodies at 1/10,000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-GSK3 alpha antibody [EP793Y] (ab40870)

Predicted band size: 51 kDa

false

• WB

Unknown

Western blot - Anti-GSK3 alpha antibody [EP793Y] (AB40870)

All lanes:

Western blot - Anti-GSK3 alpha antibody [EP793Y] (ab40870) at 1/5000 dilution

All lanes:

10ug HeLa cell lysate.

Predicted band size: 51 kDa

Observed band size: 51 kDa

false