Mouse naïve T cells stimulated in vitro as indicated above were lysed in the presence of protease/phosphatase inhibitor cocktail (catalog no. 5872; Cell Signaling Technology, Danvers, MA) for 30 min on ice. The lysates were centrifuged at 12000 × g for 30 min, 4 °C and the supernatants were subjected to standard SDS-PAGE followed by transferring to PVDF membranes for detection of STAT, T-bet or GATA-3 proteins. Rabbit anti-STAT1 (catalog no. 9172S), anti-phospho-(Tyr701) STAT 1 (catalog no. 9167S), anti-STAT4 (catalog no. 2653S), and anti-phospho-(Tyr693) STAT 4 (catalog no. 4134S) were from Cell Signaling Technology. Rabbit anti-STAT6 (catalog no. ab32520) and anti-phospho-(Tyr641) STAT 6 (catalog no. ab54461) were from Abcam. Anti-T-bet mAb 4B10 (catalog no. sc-21749) and anti-GATA-3 HG3-31 (catalog no. sc-268) were from Santa Cruz (Dallas, TX, USA).
Rabbit anti stat6
Manufactured by Abcam
Sourced in United States
Rabbit anti-STAT6 is a primary antibody that recognizes the STAT6 protein. STAT6 is a transcription factor that plays a role in cytokine signaling pathways. This antibody can be used for various applications, such as Western blotting and immunohistochemistry, to detect and study the STAT6 protein.
Automatically generated - may contain errors
Lab products found in correlation
Rabbit anti stat1, by Cell Signaling Technology (1 mentions)
Anti phospho tyr701 stat 1, by Cell Signaling Technology (1 mentions)
Anti stat4, by Cell Signaling Technology (1 mentions)
Anti gata3 hg3 31, by Santa Cruz Biotechnology (1 mentions)
Protease phosphatase inhibitor cocktail, by Cell Signaling Technology (1 mentions)
Total protein extraction kit, by Keygen Biotech (1 mentions)
Rabbit anti gapdh, by Bioworld Technology (1 mentions)
Polyvinylidene difluoride membranes, by BD (1 mentions)
Enhanced chemiluminescence, by BD (1 mentions)
Edta free protease inhibitor, by Roche (1 mentions)
Pierce ecl western blotting substrate, by Thermo Fisher Scientific (1 mentions)
Cell lysis buffer, by Beyotime (1 mentions)
Rabbit anti nrp1, by Abcam (1 mentions)
Pierce bca protein assay kit, by Thermo Fisher Scientific (1 mentions)
Ai680, by GE Healthcare (1 mentions)
Horseradish peroxidase hrp conjugated goat anti rabbit immunoglobulin g igg, by Proteintech (1 mentions)
Rabbit anti tubulin, by Proteintech (1 mentions)
3 protocols using rabbit anti stat6
1
Immunoblotting Analysis of Murine Naïve T Cells
2
Quantification of STAT6 and p-STAT6 Proteins
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Total protein was collected with Total Protein Extraction Kit (KeyGen Biotech. Co. Ltd., Nanjing, China). Proteins (30 mg) were separated on a 10% sodium dodecyl sulfate-polyacrylamide gel, and transferred electrophoretically onto polyvinylidene difluoride membranes (BD Biosciences). The membranes were blocked in 5% skimmed milk for 1 h, and then incubated for 2 h with rabbit anti-STAT6 (1:1,000; Abcam, Cambridge, MA, USA), rabbit anti-phosphorylated (p)-STAT6 (1:1,000; Cell Signaling Technology, Inc., Danvers, MA, USA) or rabbit anti-GAPDH (1:1,000; Bioworld, Technology, Inc., St. Louis Park, MN, USA), which was used as loading control. Following incubation with goat anti-rabbit secondary antibody (Bioworld, Technology, Inc.), the signals derived from the antibody-antigen binding reaction were detected by enhanced chemiluminescence (BD Biosciences).
3
Protein Analysis of HUVEC and Tumor Tissue
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HUVECs were washed twice with ice-cold PBS and lysed with cell lysis buffer (Beyotime Biotechnology) containing an EDTA-free protease inhibitor (Roche). Tumor tissue was incubated with ice-cold cell lysis buffer containing protease inhibitor and disrupted with Tissuelyser-24. Protein concentration was quantified using the Pierce™ BCA Protein Assay Kit (Thermo Fisher Scientific, Waltham, MA, USA). Western blotting was performed as previously described (23 (link)). Blots were incubated with Pierce ™ ECL Western Blotting Substrate (Thermo Fisher), and detected by AI 680 (General Electric Company, Boston, MA) Antibodies used for western blotting included rabbit anti-phospho-STAT6 (Tyr-641, 1:1000, Cell Signaling Technology); rabbit anti-STAT6 (1:1000, Abcam); rabbit anti-NRP1 (1:1000, Affinity); rabbit anti-NRP1 (1:1000, Abcam); and rabbit anti-Tubulin (1:5000, Proteintech). The secondary antibody was horseradish peroxidase (HRP)-conjugated goat anti-rabbit immunoglobulin G (IgG) (1:8000, Proteintech).
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