Tissues or cells were mechanically homogenized in ice-cold lysis buffer and centrifuged for 10 minutes at 4 °C and 14,000 g for tissues or 8,000 g for cells. The resulting supernatant fraction was separated by SDS-PAGE and immunoblotted with antibodies against phospho-AKT (S473; 1:1000), total AKT (1:1000), total JAK2 (1:1000), tubulin (1:1000), phospho-STAT3 (Y705, 1:2000), total STAT3 (1:1000), phospho-STAT5 (Y694, 1:1000), total STAT5 (1:1000), phospho-NF-κB p65 (Y694, 1:1000), total NF-κB p65 (1:1000), phospho-JNK (T183/Y185, 1:500, Santa Cruz Biotechnology) or total JNK1 (1:500, Santa Cruz Biotechnology) or GAPDH (1:5000). Antibodies were purchased from Cell Signaling Technology unless otherwise stated. Protein band intensity was quantified by ImageJ software.
Phospho stat5
Manufactured by Santa Cruz Biotechnology
Sourced in United States
Phospho-STAT5 is a lab equipment product that detects the phosphorylated form of the signal transducer and activator of transcription 5 (STAT5) protein. STAT5 is a critical transcription factor that plays a role in cellular processes such as proliferation, differentiation, and survival. The Phospho-STAT5 product allows for the identification and quantification of the activated, phosphorylated form of STAT5.
Automatically generated - may contain errors
Lab products found in correlation
Phospho nf κb p65, by Santa Cruz Biotechnology (1 mentions)
Phospho stat3, by Santa Cruz Biotechnology (1 mentions)
Phospho akt, by Santa Cruz Biotechnology (1 mentions)
Total nf κb p65, by Santa Cruz Biotechnology (1 mentions)
Gapdh, by Santa Cruz Biotechnology (1 mentions)
Horseradish peroxidase hrp conjugated goat anti rabbit and anti mouse antibodies, by Merck Group (1 mentions)
Total jak2, by Cell Signaling Technology (1 mentions)
Lipofectamine plus, by Thermo Fisher Scientific (1 mentions)
Phospho jak2, by Cell Signaling Technology (1 mentions)
Pcdna3, by Thermo Fisher Scientific (1 mentions)
2 protocols using phospho stat5
1
Western Blot Analysis of Signaling Proteins
2
Signaling Pathway Profiling in GH-Stimulated Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Phospho-JAK2 and total JAK2 were obtained from Cell Signaling Technology (USA). Phospho-STAT5 and total STAT5 were obtained from Santa Cruz (USA). Antiphosphotyrosine antibody (clone 4G10) was purchased from Upstate Biotechnology. Horseradish peroxidase (HRP)-conjugated goat anti-rabbit and anti-mouse antibodies and recombinant GH were purchased from Sigma (Sigma–Aldrich Company, St. Louis, MO, USA). 125I-GH was prepared using chloramine T according to published procedures, and the specific activity of GH was 70 to 130 μCi/μg protein. Lipofectamine Plus and pCDNA3.1 were obtained from Invitrogen (Invitrogen, Carlsbad, CA, USA). The ImmunoPure Fab Preparation kit, Cell Lysis Buffer, IP kit, Enhanced chemiluminescence (ECL) and the BCA kit were purchased from pierce (Pierce Biotechnology Inc, Rockford, IL, USA). Porcine GH receptor cDNAs were obtained from Takara Biotechnology Co., Ltd (Dalian, China). The pre-stained molecular weight standards and all tissue reagents were obtained from Gibco (Gibco, Grand Island, NY, USA). The porcine IGF-1 enzyme-linked immunosorbent assay (ELISA) kit was purchased from Hua Yi Medical and Biological Laboratories Co., Ltd (Changchun, China). Unless stated otherwise, all other reagents were from Sigma-Aldrich (USA).
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