Anti-STAT3 antibody [9D8]

11 product images

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  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) (Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) ab150113) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG (Mouse IgG - Isotype Control 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.

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  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

  Developed using the ECL technique.

  Predicted band size: 88 kDa

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  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 (Goat Anti-Mouse IgG H&L (DyLight® 488) preadsorbed ab96879) at 1/400 dilution. Nuclei (blue) were stained with Hoechst 33342 dye.

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  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

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  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

  Developed using the ECL technique.

  Predicted band size: 88 kDa

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  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.

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  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

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  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

  Developed using the ECL technique.

  Predicted band size: 88 kDa

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  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.

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  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.

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  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.