Total cellular protein was isolated from the cells after various treatments. For Western blots, a previously described procedure was applied [54 (link)]. The following primary antibodies were used: Acetyl Histone H3, HDAC1, HDAC8, PPARγ, cyclin D1, CDK6, p-Ser473 Akt, Akt, p-Ser2448 mTOR, mTOR, p-Ser139 H2AX, H2AX, Bax, Mcl-1, PARP, procaspase-8, cleaved caspase-9, LC3B, and Atg5 were purchased from Cell Signaling Technologies (Beverly, MA, USA); β-actin, Sigma-Aldrich (St. Louis, MO, USA). The secondary antibodies were purchased from Santa Cruz Biotechnology. The enhanced chemiluminescence (ECL) system for detection of immunoblotted proteins was from GE Healthcare Bioscience (Piscataway, NJ, USA). Then, the protein was visualized by FUSION SOLO S (VILBER, Deutschland, Germany).
Acetyl histone h3
Manufactured by Cell Signaling Technology
Sourced in United States
Acetyl-histone H3 is a lab equipment product from Cell Signaling Technology. It is a modified histone H3 protein that contains an acetyl group on one or more lysine residues. Histones are proteins that package and organize DNA in the cell nucleus. The acetylation of histone H3 is a post-translational modification that can alter chromatin structure and gene expression.
Automatically generated - may contain errors
Lab products found in correlation
Histone h3, by Cell Signaling Technology (9 mentions)
Hdac1, by Cell Signaling Technology (5 mentions)
β actin, by Merck Group (4 mentions)
Acetyl histone h4, by Cell Signaling Technology (4 mentions)
β catenin, by Cell Signaling Technology (3 mentions)
E cadherin, by Cell Signaling Technology (3 mentions)
Bcl2 associated x (bax), by Cell Signaling Technology (2 mentions)
Cleaved caspase 9, by Cell Signaling Technology (2 mentions)
Phospho jnk, by Cell Signaling Technology (2 mentions)
Phospho p65, by Cell Signaling Technology (2 mentions)
Taq polymerase, by Takara Bio (2 mentions)
Protein g agarose, by Merck Group (2 mentions)
Antibody to p53, by Santa Cruz Biotechnology (2 mentions)
Doxycycline, by Takara Bio (2 mentions)
Tet system approved fbs, by Takara Bio (2 mentions)
Tri methyl histone h3 lys27, by Cell Signaling Technology (2 mentions)
Histone h2b, by Cell Signaling Technology (2 mentions)
Antibodies to yy1ap1 hpa006986, by Merck Group (2 mentions)
Antibody to epcam mab960, by R&D Systems (2 mentions)
Ub histone h2b, by Cell Signaling Technology (2 mentions)
30 protocols using acetyl histone h3
1
Western Blot Analysis of Protein Signaling
2
Evaluating Chemical Compounds for Biological Activity
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VOR, and NaB and puromycin were purchased from Cayman Chemical (Ann Arbor, MI, USA) and Wako Pure Chemical (Osaka, Japan), respectively. BML-210, GI254023X, sodium DL-lactate (NaL), and BMS-303141 were purchased from Sigma-Aldrich (St. Louis, MO, USA). BEL, ENT, MOC, PNB, and RES were purchased from Selleck Chemicals (Houston, TX, USA), and ROM was purchased from Abcam (Cambridge, United Kingdom). Antibodies to histone H3 and acetyl-histone H3 were purchased from Cell Signaling Technology (Danvers, MA, USA). IL-15 and Cell Counting Kit-8 were purchased from R&D Systems (Minneapolis, MN, USA) and Dojindo (Kumamoto, Japan), respectively. Huh7 and HepG2 cells, as well as PLC/PRF/5 and the NK cell line NK92MI were obtained from American Type Culture Collection (Manassas, VA, USA), and were cultured according to the supplier’s protocols. The cell lines were authenticated by the short tandem repeat method (Bex, Tokyo, Japan) in January 2016. PXB cells were purchased from Phoenix Bio (Hiroshima, Japan).
3
SAHA Treatment Effects on Cellular Signaling
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After 24 and 48 h of treatment with various concentrations of SAHA, cell pellets were lysed directly in SDS sample buffer (50 mM Tris-HCl [pH 6.8], 2% SDS, 10% glycerol, 6% 2-mercaptoethanol and 0.0025% bromophenol blue). Cell lysates were separated on 10% acrylamide gels by SDS-PAGE, transferred to PVDF membranes, and immunoblotted with Abs. Abs were used against acetyl-histone H3 (Cell Signaling, Boston, MA, USA), poly (ADP-ribose) polymerase (PARP; Sigma, St. Louis, MO, USA), latent membrane protein (LMP) 1 (S12; BD Biosciences, San Jose, CA, USA),30 (link) EBV nuclear antigen (EBNA) 131 (link) and β-actin (Sigma).
4
ChIP Assay for Histone H3 Acetylation
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ChIP assays were conducted using a ChIP assay kit (Invitrogen) according to the manufacturer's instructions. Briefly, chromatin from cross-linked cells was sheared by sonication and incubated overnight with specific antibodies (Acetyl-Histone H3, Cell Signaling Technology) followed by incubation with protein G-Sepharose saturated with salmon sperm DNA. Precipitated DNA and input DNA were analyzed using qPCR, and the results were presented as normalization to the input DNA. The primers for the human interleukin (IL)-6 promoter were: 5′- AGACATGCCAAAGTGCTGAGT-3′ and 5′-GGCTAGCGCTAAGAAG CAGA-3′.
5
Western blot analysis of cellular signaling
6
Molecular Investigations in Neuroscience
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Agarose, PIPES, Potassium hydroxide, Tween® 20, 2-mercaptoethanol, Trypsin, MPH, ATX and VPA were purchased from Sigma (St. Louis, MO, USA). ECLTM Western blotting detection reagents were obtained from Amersham Life Science (Arlington Heights, IL, USA). Trizol and SuperScriptTM II Reverse Transcriptase were purchased from Invitrogen (Carlsbad, CA, USA). DNase Ι was purchased from Roche (Mannheim, Germany). Protein G Agarose was obtained from Millipore Corporation (Billerica, MA, USA). Taq polymerase and dNTP were obtained from Takara (Shiga, Japan). Protease inhibitor cocktail was obtained from Calbiochem (La Jolla, CA, USA). Chelex 100 was obtained from BioRad (Hercules, CA, USA). Antibodies were purchased from the following companies: anti-β-actin from Sigma (St. Louis, MO, USA), Histone H3, Acetyl-Histone H3 and HDAC1 antibody from Cell signaling (Boston, MA, USA), DAT, NET and peroxidase-conjugated secondary antibody from Santa Cruz biotechnology (Dallas, TX, USA).
7
Antibodies and Reagents for Cell Analysis
8
Quantifying Fibroblast Activation Markers
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KFBs were seeded into each well of 6-well flat bottom plates and cultured in
culture medium with 10% FBS. After 24 h, FAs were added and the plates were
incubated for 48 h. After preparation of the KFBs in 1.5 mL tubes, they were
suspended to 100 µL of Pro-Prep (iNtRON, Gyeonggi-do, Korea), according
to the manufacturer's instructions. Five microliters of the cell supernatants
was used to measure the protein concentration, using Lowry's method (RC DC
Protein Assay Kit, Bio-Rad, Hercules, CA). Western blotting was performed as
described previously, using primary antibodies against α-SMA (1:400;
Sigma), acetyl-histone H3 (1:1000; Cell Signaling Technology Inc., Danvers, CO),
PPARγ (1:800; Santa Cruz Biotechnology Inc., Santa Cruz, CA), GAPDH
(1:40000; Sigma), and appropriate horseradish peroxidase-conjugated secondary
antibody. 27 (link) Densitometric
results were analyzed using the Image J software (National Institutes of Health,
Bethesda, MD).
culture medium with 10% FBS. After 24 h, FAs were added and the plates were
incubated for 48 h. After preparation of the KFBs in 1.5 mL tubes, they were
suspended to 100 µL of Pro-Prep (iNtRON, Gyeonggi-do, Korea), according
to the manufacturer's instructions. Five microliters of the cell supernatants
was used to measure the protein concentration, using Lowry's method (RC DC
Protein Assay Kit, Bio-Rad, Hercules, CA). Western blotting was performed as
described previously, using primary antibodies against α-SMA (1:400;
Sigma), acetyl-histone H3 (1:1000; Cell Signaling Technology Inc., Danvers, CO),
PPARγ (1:800; Santa Cruz Biotechnology Inc., Santa Cruz, CA), GAPDH
(1:40000; Sigma), and appropriate horseradish peroxidase-conjugated secondary
antibody. 27 (link) Densitometric
results were analyzed using the Image J software (National Institutes of Health,
Bethesda, MD).
9
Histone and Tubulin Acetylation Assay
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For analyzing acetylation status of histone 3 and tubulin, NIH3T3 cells were grown to confluence in 6-well plates. After 48hrs of treatment with indicated chemicals, cells were lysed in RIPA buffer or 1% NP40/phosphate buffered saline/protease inhibitors (SIGMA, #S8820), the lysate cleared using a microcentrifuge, then 6 × sample loading buffer (Bioland Scientific) added to the supernatant. Proteins were separated on SDS-PAGE (BioRad Criterion TGX Precast Gels). Antibodies used for analyzing the blots were: acetylated tubulin (SIGMA, #T6793), tubulin, acetyl-histone H3 (Lys23), and histone H3 (Cell Signaling Technology, #2125 S, #8848 and #9717, respectively). Chemiluminescence was detected using a Li-COR Odyssey Imaging System.
10
Protein Expression Profiling in Cell Lysates
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