IHC and immunoblotting were performed as we previously described 10 (link), 68 (link). Antibodies for IHC included p-STAT3 (Y705) antibody (Cell Signaling; #9145; 1:50), and rabbit polyclonal GLI1 and tGLI1 antibodies that we developed and validated 10 (link), 26 , 68 (link). Antibodies for immunoblotting included p-STAT3 (Y705) antibody (Cell Signaling, 1:1,000), STAT3 rabbit polyclonal antibody (Santa Cruz; C20; 1:1,000), α-tubulin and β-actin mouse monoclonal antibodies (Sigma), and rabbit polyclonal GLI1/tGLI1 antibody (Cell Signaling, 1:1,000). For IP, cells were transfected with flag-tagged GLI1, tGLI1 or STAT3 mutants, stimulated with 100 ng/ml EGF and 100 ng/ml SHH for 2 hrs, washed, fractionated, and precleared with protein G-agarose (Sigma; A2220). Precleared lysates were incubated with anti-flag M2 affinity gel or mouse IgG at 4°C overnight with gentle agitation. Pellets were collected, washed and subjected to SDS-PAGE and WB analysis. STAT3 IP was performed using antibodies from Santa Cruz (C-20). GLI1 and tGLI1 immunoprecipitation was performed using a GLI1 antibody (Santa Cruz, H-300) that recognizes the COOH-terminal end and, therefore, pulls down GLI1 and tGLI1.
Stat3 rabbit polyclonal antibody
Manufactured by Santa Cruz Biotechnology
The STAT3 rabbit polyclonal antibody is a laboratory tool used to detect the presence and quantify the levels of the STAT3 protein in biological samples. STAT3 is a key transcription factor involved in various cellular processes. This antibody can be utilized in techniques such as Western blotting, immunohistochemistry, and immunoprecipitation to study the expression and localization of STAT3 in cells and tissues.
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2 protocols using stat3 rabbit polyclonal antibody
1
Characterizing STAT3 and GLI1 Interactions
2
Characterizing STAT3 and GLI1 Interactions
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IHC and immunoblotting were performed as we previously described 10 (link), 68 (link). Antibodies for IHC included p-STAT3 (Y705) antibody (Cell Signaling; #9145; 1:50), and rabbit polyclonal GLI1 and tGLI1 antibodies that we developed and validated 10 (link), 26 , 68 (link). Antibodies for immunoblotting included p-STAT3 (Y705) antibody (Cell Signaling, 1:1,000), STAT3 rabbit polyclonal antibody (Santa Cruz; C20; 1:1,000), α-tubulin and β-actin mouse monoclonal antibodies (Sigma), and rabbit polyclonal GLI1/tGLI1 antibody (Cell Signaling, 1:1,000). For IP, cells were transfected with flag-tagged GLI1, tGLI1 or STAT3 mutants, stimulated with 100 ng/ml EGF and 100 ng/ml SHH for 2 hrs, washed, fractionated, and precleared with protein G-agarose (Sigma; A2220). Precleared lysates were incubated with anti-flag M2 affinity gel or mouse IgG at 4°C overnight with gentle agitation. Pellets were collected, washed and subjected to SDS-PAGE and WB analysis. STAT3 IP was performed using antibodies from Santa Cruz (C-20). GLI1 and tGLI1 immunoprecipitation was performed using a GLI1 antibody (Santa Cruz, H-300) that recognizes the COOH-terminal end and, therefore, pulls down GLI1 and tGLI1.
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