Anti-STAT3 antibody [9D8]

• WB

Unknown

Western blot - Anti-STAT3 antibody [9D8] (AB119352)

4-20% Tris-HCl polyacrylamide gel

All lanes:

Western blot - Anti-STAT3 antibody [9D8] (ab119352) at 1/1000 dilution

Lane 1:

U2OS lysate from non-targeting control at 25 µg

Lane 2:

U2OS lysate from STAT3 SMART pool siRNA transfected at 25 µg

Secondary

All lanes:

goat anti-mouse-HRP at 1/20000 dilution

Predicted band size: 88 kDa

true

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT3 antibody [9D8] (AB119352)

ab119352, at 1/1600 dilution, staining STAT3 in Human brain tumor tissue by Immunohistochemistry. Detection was performed using a goat anti-mouse HRP secondary antibody followed by colorimetric detection using DAB substrate. Tissues were counterstained with hematoxylin.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT3 antibody [9D8] (AB119352)

ab119352, at 1/1600 dilution, staining STAT3 in Human stomach cancer tissue by Immunohistochemistry. Detection was performed using a goat anti-mouse HRP secondary antibody followed by colorimetric detection using DAB substrate. Tissues were counterstained with hematoxylin.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-STAT3 antibody [9D8] (AB119352)

Immunofluorescent analysis of STAT3 using ab119352 at 1/100 dilution (shown in green) in HeLa cells (formalin fixed cells permeabilized with 0.1% Triton X-100) using a DyLight^® 488 goat-anti-mouse secondary antibody (ab96879) at 1/400 dilution. Nuclei (blue) were stained with Hoechst 33342 dye.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT3 antibody [9D8] (AB119352)

ab119352, at 1/1600 dilution, staining STAT3 in Human pancreas tissue by Immunohistochemistry. Detection was performed using a goat anti-mouse HRP secondary antibody followed by colorimetric detection using DAB substrate. Tissues were counterstained with hematoxylin.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT3 antibody [9D8] (AB119352)

ab119352, at 1/1600 dilution, staining STAT3 in Human colon cancer tissue by Immunohistochemistry. Detection was performed using a goat anti-mouse HRP secondary antibody followed by colorimetric detection using DAB substrate. Tissues were counterstained with hematoxylin.

• Flow Cyt

Lab

Flow Cytometry - Anti-STAT3 antibody [9D8] (AB119352)

Overlay histogram showing HeLa cells stained with ab119352 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween 20 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 (ab119352, 0.1μg/1x10⁶ cells) for 30 min at 22°C. The secondary antibody used was Alexa Fluor^® 488 goat anti-mouse IgG (H&L) (ab150113) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG (ab37355, 0.1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control.

Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

• IP

Unknown

Immunoprecipitation - Anti-STAT3 antibody [9D8] (AB119352)

Immunoprecipitation of STAT3 was performed on HepG2 cells. The antigen : antibody complex was formed by incubating 750µg whole cell lysate with 2µg of ab119352 overnight at 4°C. The immune-complex was captured, washed extensively and proteins eluted with 5X Reducing Sample Loading Dye. Samples were resolved on a 4-20% Tris-HCl polyacrylamide gel. Proteins were transferred to PVDF membrane and blocked with 5% Milk/TBS-0.1% Tween for at least 1 hour. Membranes were then probed with ab119352 at a dilution of 1/5000 overnight at 4°C. Membranes were washed in TBST and probed with IP detection reagent at a dilution of 1/2000 for at least one hour. Membranes were washed and chemiluminescent detection was performed.

All lanes:

Immunoprecipitation - Anti-STAT3 antibody [9D8] (ab119352)

Predicted band size: 88 kDa

false

• WB

Unknown

Western blot - Anti-STAT3 antibody [9D8] (AB119352)

4-20% Tris-HCl polyacrylamide gel

All lanes:

Western blot - Anti-STAT3 antibody [9D8] (ab119352) at 1/5000 dilution

Lane 1:

HepG2 cell lysate at 25 µg

Lane 2:

293T cell lysate at 25 µg

Lane 3:

Jurkat cell lysate at 25 µg

Lane 4:

A431 cell lysate at 25 µg

Lane 5:

U2OS cell lysate at 25 µg

Lane 6:

MCF7 cell lysate at 25 µg

Lane 7:

A549 cell lysate at 25 µg

Lane 8:

K562 cell lysate at 25 µg

Lane 9:

COS7 cell lysate at 25 µg

Lane 10:

NIH3T3 cell lysate at 25 µg

Lane 11:

C2C12 cell lysate at 25 µg

Lane 12:

NRK cell lysate at 25 µg

Secondary

All lanes:

goat anti-mouse-HRP at 1/20000 dilution

Predicted band size: 88 kDa

true

• WB

Unknown

Western blot - Anti-STAT3 antibody [9D8] (AB119352)

4-20% Tris-HCl polyacrylamide gel

All lanes:

Western blot - Anti-STAT3 antibody [9D8] (ab119352) at 1/1000 dilution

All lanes:

HepG2 total lysate at 25 µg

Secondary

All lanes:

goat anti-mouse-HRP at 1/20000 dilution

Predicted band size: 88 kDa

true

• WB

Supplier Data

Western blot - Anti-STAT3 antibody [9D8] (AB119352)

All lanes:

Western blot - Anti-STAT3 antibody [9D8] (ab119352) at 1/5000 dilution

Lane 1:

Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate at 30 µg

Lane 2:

A-431 (Human epidermoid carcinoma cell line) whole cell lysate at 30 µg

Lane 3:

Hep G2 (Human liver hepatocellular carcinoma cell line) whole cell lysate at 30 µg

Lane 4:

HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate at 30 µg

Lane 5:

PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate at 30 µg

Lane 6:

NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate at 30 µg

Lane 7:

A549 (Human lung carcinoma cell line) whole cell lysate at 30 µg

Lane 8:

Mouse lung tissue lysate at 30 µg

Predicted band size: 88 kDa

Observed band size: 80 kDa

false