Cells were lysed in LDS sample buffer, 50 μg total protein was separated on NuPAGE Novex 4–12% Bis-Tris Protein Gels (ThermoFisher Scientific) and transferred to 0.2 μm nitrocellulose membranes (GE Healthcare, Vienna, Austria). The following antibodies were used: anti-MAO-A rabbit Mab (EPR7101, 1:2000) (Abcam), anti-E-cadherin (Cat:610181, 1:500) (Becton Dickinson, Heidelberg, Germany), Vimentin (SC6260, 1:500) (Cruz Biotechnology, Santa Cruz, CA), anti-GAPDH (MAB374, 1:50.000) (Millipore, Vienna, Austria), anti cPARP (G7341, 1:500) (Sigma), and anti-p21 (2946 S, 1:500) (Cell Signaling, Frankfurt, Germany). Specificity of the used MAO-A antibody was tested and confirmed using protein of PF179TCAF cells after treatment with either 100 nM Dex, or a combination of 100 nM Dex and 6 µM RU486 for 3 days and using protein of PC3 cells overexpressing MAO-A. Furthermore, reduced MAO-A protein was confirmed after transient MAO-A knockdown for 3 days in LNCaPabl cells utilizing ON-TARGET plus Human MAO-A siRNA SMARTpool (L-009369-00-0005), siControl On-Target plus Non-targeting pool (D-001810-10-05) and Lipofectamine2000 (ThermoFisher Scientific) according to the manufacturer’s protocols.
Anti p21 2946 s
Manufactured by Cell Signaling Technology
Anti-p21 (2946S) is a primary antibody product offered by Cell Signaling Technology. It is designed for the detection of p21 protein, which is a cyclin-dependent kinase inhibitor involved in cell cycle regulation. The antibody can be used in various applications such as Western blotting, immunoprecipitation, and immunohistochemistry to study the expression and localization of p21 in biological samples.
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Lab products found in correlation
Lipofectamine 2000, by Thermo Fisher Scientific (2 mentions)
Nupage novex 4 12 bis tris protein gel, by Thermo Fisher Scientific (1 mentions)
0.2 μm nitrocellulose membrane, by GE Healthcare (1 mentions)
Anti e cadherin, by BD (1 mentions)
Anti gapdh mab374, by Merck Group (1 mentions)
Cell lysis buffer, by Cell Signaling Technology (1 mentions)
Anti p ser15 p53, by Cell Signaling Technology (1 mentions)
Anti usp7 a300 033a, by Fortis Life Sciences (1 mentions)
Phosstop phosphatase inhibitor, by Roche (1 mentions)
2 protocols using anti p21 2946 s
1
Western Blot Analysis of Protein Expression
2
Western Blot Analysis of p53 Pathway
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Whole-cell lysates were extracted in cell lysis buffer (Cell Signaling) supplemented with EDTA-free protease inhibitors and PhosSTOP phosphatase inhibitors (Roche). Western immunoblotting was performed using standard techniques. Primary antibodies used included anti-MDM2 (ab178938; Abcam), anti-MDM2 (86934; Cell Signaling), anti-MDM4 (A300-287A; Bethyl Laboratories), anti-p53 (2527S; Cell Signaling), anti-p21 (2946S; Cell Signaling), anti-Vinculin (18058; Abcam), anti-Wip1 (A300-664A; Bethyl Laboratories), anti-pSer15-p53 (9284; Cell Signaling), anti-USP7 (A300-033A; Bethyl Laboratories), and anti-Tubulin (cp06; CalBiochem).
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