Anti-STAT3 antibody [84/Stat3]

• WB

Supplier Data

Western blot - Anti-STAT3 antibody [84/Stat3] (AB280212)

Lysates should be made freshly and used in WB immediately to minimize protein degradation.

Blocking/Dilution buffer : 5% NFDM/TBST.

Exposure times : Lane 1/4-6 : 37 secs;

Lanes 2-3 : 15 secs.

All lanes:

Western blot - Anti-STAT3 antibody [84/Stat3] (ab280212) at 1/1000 dilution

Lane 1:

RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage), whole cell lysate at 10 µg

Lane 2:

THP-1 (human monocytic leukemia monocyte), whole cell lysate at 10 µg

Lane 3:

Jurkat (human T cell leukemia T lymphocyte), whole cell lysate at 10 µg

Lane 4:

HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate at 10 µg

Lane 5:

NIH/3T3 (mouse embryonic fibroblast), whole cell lysate at 10 µg

Lane 6:

PC-12 (rat adrenal gland pheochromocytoma), whole cell lysate at 10 µg

Secondary

All lanes:

Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/10000 dilution

Predicted band size: 88 kDa

false

• WB

Supplier Data

Western blot - Anti-STAT3 antibody [84/Stat3] (AB280212)

Lanes 1 - 4 : Merged signal (red and green). Green - ab280212 observed at 88 kDa. Red - loading control ab181602(Rabbit monoclonal [EPR16891] to GAPDH) observed at 36 kDa.

Lanes 1-2 : ab280212 Anti-Stat3 antibody was shown to react with in HeLa cells in Western blot. Loss of signal was observed when Stat3 knockout sample was used. Wild-type and Stat3 knockout samples were subjected to SDS-PAGE.

ab280212 and Anti-GAPDH antibody [EPR16891] (ab181602) were incubated at 4℃ overnight at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 680CW) (ab216777) and Goat anti-Mouse IgG H&L (IRDye® 800RD) (ab216772) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging.

Blocking/Dilution buffer : Intercept® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBS.

All lanes:

Western blot - Anti-STAT3 antibody [84/Stat3] (ab280212) at 1/1000 dilution

Lane 1:

Wild-type HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate at 10 µg

Lane 2:

Stat3 knockout HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate at 10 µg

Lane 3:

K562 (human chronic myelogenous leukemia lymphoblast), whole cell lysate at 10 µg

Lane 4:

Jurkat (human T cell leukemia T lymphocyte), whole cell lysate at 10 µg

Secondary

All lanes:

Goat Anti-Mouse IgG H&L (IRDye® 800CW) (<a href='/en-us/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-800cw-preadsorbed-ab216772'>ab216772</a>) and Goat Anti-Rabbit IgG H&L (IRDye® 680RD) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-680rd-preadsorbed-ab216777'>ab216777</a>) at 1/10000 dilution

Predicted band size: 88 kDa

false