Cells were washed with phosphate-buffered saline and scraped into chilled lysis buffer (8 M urea, 1 M thiourea, 0.5% CHAPS (3-((3-cholamidopropyl)dimethylammonio)-1-propanesulfonate), 50 μM dithiothreitol and 24 μM spermine). Lysates were assayed for protein using Bradford reagent (Pierce, Rockford, IL, USA), resolved by denaturing sodium dodecyl sulfate–polyacrylamide gel electrophoresis, and then electroblotted onto polyvinylidene difluoride membranes (Millipore, Billerica, MA, USA). Proteins were detected with antibodies to: pan-MAGEA (clone 6C1; Santa Cruz, Dallas, TX, USA; sc-20034), MAGEA2 (sc-130164), MAGEA3 (sc-130809), p53 (DO1, sc-126), acetylated(Ac)-p53 (Cell Signaling, Beverly, MA, USA; no. 2525), lamin A/C (Cell Signaling; no. 4477), p21cip (Cell Signaling; no. 2946), ERα (sc-56833), phospho-ERα (Ser 118) (Cell Signaling, no. 2511), p42/44 MAPK (Cell Signaling; no. 9102), phospho-p42/44 MAPK (Cell Signaling; no. 4370), actin (sc-130301) or Hsc 70 (sc-65521), and visualized with ECL chemiluminescence reagent (Amersham, Buckinghamshire, UK).
Acetylated ac p53
Manufactured by Cell Signaling Technology
Sourced in United States
Acetylated(Ac)-p53 is a product from Cell Signaling Technology that detects acetylated p53 protein. p53 is a tumor suppressor protein that is post-translationally modified by acetylation, which can influence its activity and stability.
Automatically generated - may contain errors
Lab products found in correlation
P21cip, by Cell Signaling Technology (1 mentions)
Polyvinylidene difluoride membrane, by Merck Group (1 mentions)
P42 44 mapk, by Cell Signaling Technology (1 mentions)
Phospho p42 44 mapk, by Cell Signaling Technology (1 mentions)
P53 do 1, by Cell Signaling Technology (1 mentions)
Lamin a c, by Cell Signaling Technology (1 mentions)
Phospho erαser118, by Cell Signaling Technology (1 mentions)
Enhanced chemiluminescence, by Merck Group (1 mentions)
Anti mouse and anti rabbit secondary antibodies, by Cell Signaling Technology (1 mentions)
Antibodies against parp 1, by Cell Signaling Technology (1 mentions)
Anti β actin antibody, by Cell Signaling Technology (1 mentions)
Pvdf membrane, by Merck Group (1 mentions)
2 protocols using acetylated ac p53
1
Protein Quantification and Detection
2
Protein Extraction and Western Blot Analysis
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Cellular proteins were extracted using radioimmunoprecipitation assay buffer, and protein concentrations were measured using the bicinchoninic acid method. Approximately 30 μg of protein per well was loaded on 8 or 10% gels for sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Proteins were transferred to polyvinylidene difluoride (PVDF) membranes (Millipore), and sequentially detected by primary antibodies, secondary antibodies, and enhanced chemiluminescence (Millipore). Antibodies against PARP1, SIRT1, acetylated (ac)-p53, p53, and cyclin-dependent kinase inhibitor 1A (p21), as well as anti-mouse and anti-rabbit secondary antibodies, were purchased from Cell Signaling Technology. The anti-PAR antibody was purchased from Invitrogen. An anti-β-actin antibody (Cell Signaling Technology) was used as an internal control.
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