Whole-cell extracts were prepared as described previously (Liu et al. 2013 (link)). In brief, equivalent amounts of protein (50 μg) were separated by sodium dodecyl-sulfate-polyacrylamide gel electrophoresis (10 %) and transferred to nitrocellulose membrane (Bio-Rad, Hercules, CA). The membranes were subsequently incubated with the following primary antibodies: GTSE1 (Proteintech Group, Inc), AKT, phospho-AKT (Ser473), and cyclin B1 from CST (Cell Signaling Technology, Beverly, MA, USA), BCL-2 and Bax from Abcam (AbCam, Cambridge, MA, USA), and GAPDH (CST) was used as a loading control. The blots were visualized with a horseradish peroxidase-conjugated secondary antibody (Kangchen, Shanghai, China) and an enhanced chemiluminescence kit (Millipore, Bedford, MA).
Bcl 2 and bax
Manufactured by Abcam
Sourced in United States, United Kingdom
BCL-2 and Bax are proteins involved in the regulation of apoptosis, a process of programmed cell death. BCL-2 is an anti-apoptotic protein that promotes cell survival, while Bax is a pro-apoptotic protein that induces cell death. These proteins play a crucial role in the balance between cell survival and cell death.
Automatically generated - may contain errors
Lab products found in correlation
Enhanced chemiluminescence kit, by Merck Group (1 mentions)
Cyclin b1, by Cell Signaling Technology (1 mentions)
Gtse1, by Proteintech (1 mentions)
Phospho akt ser473, by Cell Signaling Technology (1 mentions)
Nitrocellulose membrane, by Bio-Rad (1 mentions)
Gapdh, by Abcam (1 mentions)
P smad2, by Abcam (1 mentions)
P smad3, by Abcam (1 mentions)
Tgf βri, by Abcam (1 mentions)
Smad2, by Abcam (1 mentions)
Anti tgf β1, by Abcam (1 mentions)
Smad3, by Abcam (1 mentions)
2 protocols using bcl 2 and bax
1
Western Blot Analysis of Cell Signaling
2
Investigating TGF-β1 Signaling Pathway
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The prostatic expressions of transforming growth factor-β1 (TGF-β1), type I TGF-β receptor (TGF-βRI), Smad2, phosphorylation-Smad2 (p-Smad2), Smad3, p-Smad3, Smad7, nuclear related factor-2 (Nrf-2), heme oxygenase-1 (HO-1), B-cell CLL/lymphoma (Bcl)-2 and Bcl-2-associated X protein (Bax) were assessed by western blot analysis. Tissue protein samples were separated by 10% SDS-polyacrylamide gel electrophoresis and then transferred to a PVDF membrane by electrophoretic transfer. The membranes were blocked for 1 h at 37˚C with 5% nonfat milk in Tris-buffered saline, and then incubated overnight at 4˚C with the primary antibodies (anti-TGF-β1, TGF-βRI, Smad2, p-Smad2, Smad3, p-Smad3, Smad7, Bcl-2 and Bax (Abcam, UK), or -Nrf-2 and HO-1 (Proteintech Group, USA)), respectively. After washed with TBST, the membranes were incubated with horseradish peroxidase-conjugated secondary antibodies in TBST with 3% nonfat milk for 1 h at room temperature. Immunoblots were developed and then the quantification of bands was determined by integrated optical density analysis using Gel-Pro Analyzer software. The result of nuclear Nrf-2 was normalized using LaminB as an internal control. The other data were normalized using β-actin as an internal control.
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