Human/Mouse/Rat STAT3 Antibody

Detection of STAT3 in HeLa Human Cell Line by Flow Cytometry.

HeLa human cell line was stained with Mouse Anti-Human/Mouse/Rat STAT3 Monoclonal Antibody (Catalog # MAB1799, filled histogram) or isotype control antibody (Catalog # MAB0041, open histogram) followed by anti-Mouse IgG PE-conjugated secondary antibody (Catalog # F0102B). To facilitate intracellular staining, cells were fixed with Flow Cytometry Fixation Buffer (Catalog # FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (Catalog # FC005). View our protocol for Staining Intracellular Molecules.

STAT3 Specificity is Shown by Immunocytochemistry in Knockout Cell Line.

STAT3 was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line treated with IFN-alpha 1, but is not detected in STAT3 knockout (KO) HeLa cell line using Mouse Anti-Human/ Mouse/Rat STAT3 Monoclonal Antibody (Catalog # MAB1799) at 1 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm and nuclei. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Western Blot Shows Human STAT3 Specificity by Using Knockout Cell Line.

Western blot shows lysates of HeLa human cervical epithelial carcinoma parental cell line, STAT1 knockout (KO) HeLa cell line, STAT2 KO HeLa cell line, STAT3 KO HeLa cell line, STAT5a KO HeLa cell line, STAT5b KO HeLa cell line, STAT6 KO HeLa cell line,. PVDF membrane was probed with 0.5 µg/mL of Mouse Anti-Human/Mouse/Rat STAT3 Monoclonal Antibody (Catalog # MAB1799) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for STAT3 at approximately 80 kDa (as indicated) in the parental HeLa cell line, but is not detectable in the STAT3 knockout HeLa cell line. GAPDH (Catalog # AF5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

STAT3 Specificity is Shown by Flow Cytometry in Knockout Cell Line.

STAT3 knockout HeLa human cervical epithelial cell line was stained with Mouse Anti-Human/Mouse STAT3 Monoclonal Antibody (Catalog # MAB1799, filled histogram) or isotype control antibody (Catalog # MAB0041, open histogram) followed by anti-Mouse IgG PE-conjugated secondary antibody (Catalog # F0102B). No staining in the STAT3 knockout HeLa cell line was observed. To facilitate intracellular staining, cells were fixed with Flow Cytometry Fixation Buffer (Catalog # FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (Catalog # FC005). View our protocol for Staining Intracellular Molecules.

Detection of Mouse STAT3 by Western Blot

Effects of kurarinone on the phosphorylation of p-STAT1 and p-STAT3 in lymph nodes. Single cell suspensions were collected from ILNs on day 42, the protein expression levels of p-STAT1, STAT1, p-STAT3, and STAT3 were measured using Western blots. (A) Representative images of Western blot and (B) Densitometric analysis for protein expressions was performed using ImageJ software. Data are presented as mean ± SEM of 6 mice from one of three experiments. (*) p < 0.05, (**) p < 0.01, (***) p < 0.001 versus vehicle-treated CIA mice group (One Way ANOVA followed by Tukey’s multiple comparison test). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/33924467), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of STAT3 by Western Blot

Galectin-3 expression induces activation of PYK2, STAT1 and GSK3 alpha/ beta signalling. Expression of 37 protein kinases in SW620 cells in response to 10 µg/ml galectin-3 or BSA for 0.5 h was assessed by Proteome Profiler Human Phospho-Kinase Array (A, Percentage changes of the kinases in cell response to galectin-3 in comparison to control are shown at the bottom panel). The presence of galectin-3 increases the phosphorylation of PYK2, GSK3 alpha/ beta, and STAT1 and decreases phosphorylation of STAT3. SW620 cells treated with 10 µg/ml galectin-3 for different times were assessed by immunoblotting using antibodies against p-PYK2, p-STAT-1, p-GSK3 alpha/ beta or p-STAT-3 (B). The blots were striped and reprobed with antibodies against PYK2, STAT-1, GSK3 alpha/ beta or STAT-3. The band density was quantified and expressed as percentages of phospho-/non-phosphorylated proteins (C). In D and E, SW620 cells were treated with 10 µg/ml galectin-3 or BSA followed by introduction of GSK3 alpha/ beta inhibitor SB 216763 (SB) or PKY2 inhibitor PF-431396 (PF) for 15 min and the levels of phosphorylated PYK2, STAT-1, GSK3 alpha/ beta or STAT-3 were analysed by immunoblotting. The blots were striped and reprobed with antibodies against PYK2, STAT-1, GSK3 alpha/ beta or STAT-3. The densities of the blots from three independent experiments were quantified and are expressed as the percentage of phosphorylated/non-phosphorylated levels of each protein. ***P < 0.001, **P < 0.01, *P < 0.05 (ANOVA). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/37055381), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of STAT3 by Western Blot

Galectin-3 expression induces activation of PYK2, STAT1 and GSK3 alpha/ beta signalling. Expression of 37 protein kinases in SW620 cells in response to 10 µg/ml galectin-3 or BSA for 0.5 h was assessed by Proteome Profiler Human Phospho-Kinase Array (A, Percentage changes of the kinases in cell response to galectin-3 in comparison to control are shown at the bottom panel). The presence of galectin-3 increases the phosphorylation of PYK2, GSK3 alpha/ beta, and STAT1 and decreases phosphorylation of STAT3. SW620 cells treated with 10 µg/ml galectin-3 for different times were assessed by immunoblotting using antibodies against p-PYK2, p-STAT-1, p-GSK3 alpha/ beta or p-STAT-3 (B). The blots were striped and reprobed with antibodies against PYK2, STAT-1, GSK3 alpha/ beta or STAT-3. The band density was quantified and expressed as percentages of phospho-/non-phosphorylated proteins (C). In D and E, SW620 cells were treated with 10 µg/ml galectin-3 or BSA followed by introduction of GSK3 alpha/ beta inhibitor SB 216763 (SB) or PKY2 inhibitor PF-431396 (PF) for 15 min and the levels of phosphorylated PYK2, STAT-1, GSK3 alpha/ beta or STAT-3 were analysed by immunoblotting. The blots were striped and reprobed with antibodies against PYK2, STAT-1, GSK3 alpha/ beta or STAT-3. The densities of the blots from three independent experiments were quantified and are expressed as the percentage of phosphorylated/non-phosphorylated levels of each protein. ***P < 0.001, **P < 0.01, *P < 0.05 (ANOVA). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/37055381), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of STAT3 by Western Blot

Aspirin modulated the JAK/p-STAT3 signaling pathway in atypical hyperplastic intestinal mucosal cells of UC mice (n = 4 for each group). (A) Western blotting images for JAK/p-STAT3 signaling pathway-associated molecules expression. (B) Statistical analysis and comparison for p-STAT3 expression. (C) Statistical analysis and comparison for STAT3 expression. (D) Statistical analysis and comparison for cyclin D1 expression. (E) Statistical analysis and comparison for SOCS3 expression. *P <.05 versus control group. uc model jak janus kinase ulcerative colitis p-stat3 phosphorylated-stat3 stat3 signal transducer and activator of transcription socs3 suppressor cytokine signaling image collected cropped by citeab from the following open publication licensed under a cc-by license. not internally tested r systems.>