RRMM-BMMNC were treated with different concentrations of LBH589, and 24 h later, total cellular proteins were extracted by using RIPA lysis buffer (Beyotime Biotechnology, Shanghai, China). The protein samples were separated by using 10% SDS-PAGE gels, and then transferred to a PVDF membrane (Millipore, Bedford, MA). After blocking with 5% non-fat milk, the membranes were then probed with antibodies against Acetyl-H4 (1: 1000, Cell Signaling Technology), PARP (1: 1000, Cell Signaling Technology), and Bcl-x (1: 1000, Cell Signaling Technology). Membranes were washed 3 times and then incubated with horseradish peroxidase (HRP)- conjugated secondary antibodies. The enhanced chemiluminescence detection system was used to visualize the immunoreactive bands.
Acetyl h4
Manufactured by Cell Signaling Technology
Sourced in United States
Acetyl-H4 is a laboratory reagent used in the detection and quantification of acetylated histone H4 proteins. It serves as a specific antibody targeting the acetylated form of histone H4, enabling researchers to study epigenetic modifications and chromatin dynamics.
Automatically generated - may contain errors
Lab products found in correlation
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (2 mentions)
Bcl x, by Cell Signaling Technology (1 mentions)
Pvdf membrane, by Merck Group (1 mentions)
Ripa lysis buffer, by Beyotime (1 mentions)
Quant it dsdna hs assay, by Thermo Fisher Scientific (1 mentions)
Sc 858, by Santa Cruz Biotechnology (1 mentions)
Sc 9001, by Santa Cruz Biotechnology (1 mentions)
Cytochrome c, by Cell Signaling Technology (1 mentions)
Suberoylanilide hydroxamic acid saha, by Merck Group (1 mentions)
Cyclin a, by Cell Signaling Technology (1 mentions)
Poly adp ribose polymerase parp, by Cell Signaling Technology (1 mentions)
2 7 dichlorofluorescein diacetate dcfh da, by Merck Group (1 mentions)
Cyclin b, by Cell Signaling Technology (1 mentions)
Cyclin d, by Cell Signaling Technology (1 mentions)
Methyl thiazolyl tetrazolium (mtt), by Thermo Fisher Scientific (1 mentions)
Bcl2 associated x (bax), by Cell Signaling Technology (1 mentions)
Bcl 2, by Cell Signaling Technology (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
β actin, by Cell Signaling Technology (1 mentions)
3 protocols using acetyl h4
1
Protein Expression Analysis of RRMM-BMMNC
2
Chromatin Immunoprecipitation Protocol for BCL6 and RNA Pol II
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ChIP was performed essentially as described [35 (link)]. Cells were fixed in 1% formaldehyde for 10 minutes, sonicated, and lysates were immunoprecipitated overnight with 2 µg of antibodies to BCL6 (sc-858) and RNA polymerase II (sc-9001) from Santa Cruz Biotechnology or acetyl H4 from Cell Signaling Technology. ChIP product was measured using Quant-iT™ dsDNA HS Assay (Invitrogen), and equal amounts of product and input were analyzed by quantitative PCR. Data are expressed as fold binding over background (using both input and genomic regions known not to be bound by BCL6, including the rhodopsin gene and a previously defined control region found within the BCL6 gene [20 (link)]). For siRNA ChIPs, 7.5×106 SK-BR-3, T-47D, or MDA-MB-468 cells were plated and ChIP was performed 72 hours after reverse transfection. ChIP product was analyzed by qPCR using the indicate primers (Supplementary Table 1 or as described [3 (link)]). OCI-Ly1 ChIP was performed essentially as above except cells were lysed in RIPA buffer (150 mM NaCl, 1% v/v Nonidet P-40, 0.5% w/v deoxycholate, 0.1% w/v SDS, 50 mM Tris pH 8, and 5 mM EDTA).
3
Characterization of SAHA and MHY4381 Effects
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Suberoylanilide hydroxamic acid (SAHA) and other chemicals were obtained from Sigma-Aldrich (St. Louis, MO, USA). MHY4381 (purity, 98.5%) was synthesized by Prof. Moon (College of Pharmacy, Pusan National University, Busan, Korea). Stock solutions (10 mM) of drugs were prepared in sterile dimethyl sulfoxide (DMSO) and stored at −20°C until use. Culture media and fetal bovine serum (FBS) were purchased from Gibco Invitrogen Corporation (Carlsbad, CA, USA). Primary antibodies against acetyl-H3, acetyl-H4, Bax, poly-ADP-ribose polymerase (PARP), Bcl-2, p53, cytochrome C, cyclin A, cyclin B, cyclin D, HDACs, caspase-3 and 9, and β-actin were purchased from Cell Signaling (Beverly, MA, USA). Antibodies against p27 and p21 and horseradish peroxidase (HRP)-conjugated secondary antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide) and DAPI (4′-6-diamidino-2-phenylindole) were purchased from Thermo Fisher Scientific, Inc (Invitrogen, MA, USA), while DCFH-DA (2′,7′-dichlo rofluorescein-diacetate) was obtained from Sigma-Aldrich.
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