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Hdac7

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HDAC7 is a histone deacetylase enzyme that regulates gene expression by removing acetyl groups from histones, leading to a more condensed chromatin structure. It plays a role in various cellular processes, including cell differentiation and proliferation.

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Lab products found in correlation

Hdac1, by Cell Signaling Technology (6 mentions) Hdac4, by Cell Signaling Technology (6 mentions) Hdac5, by Cell Signaling Technology (5 mentions) Hdac2, by Cell Signaling Technology (5 mentions) Hdac6, by Cell Signaling Technology (5 mentions) Hdac3, by Cell Signaling Technology (4 mentions) Calnexin, by Santa Cruz Biotechnology (2 mentions) Hdac6, by Santa Cruz Biotechnology (2 mentions) Nitrocellulose membrane, by Bio-Rad (2 mentions) Pan histone h3, by Abcam (2 mentions) Hnrnpl 4d11, by Abcam (2 mentions) Flag tag (flag), by Cell Signaling Technology (2 mentions) Lamin b1, by Abcam (2 mentions) H3k27ac, by Abcam (2 mentions) H3k9ac, by Abcam (2 mentions) Survivin, by Cell Signaling Technology (2 mentions) Protein inhibitor cocktail, by Merck Group (1 mentions) Anti nrf2, by Abcam (1 mentions) Anti nqo1, by Abcam (1 mentions) Anti ho 1, by Abcam (1 mentions)

10 protocols using hdac7

1

Immunoblotting of HDAC Isoforms

Cited 1 time
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Immunoblotting was performed as previously described(16 (link)). Briefly, pediatric human and neonatal rat RV homogenates were prepared and concentrations quantified as above for HDAC catalytic activity assay. Proteins were resolved by SDS-PAGE, transferred to nitrocellulose membranes (BioRad) and probed with antibodies for HDAC1 (Cell Signaling Technology, 5356), HDAC2 (Cell Signaling Technology, 5113), HDAC3 (Cell Signaling Technology, 3949), HDAC4 (Cell Signaling Technology, 5392), HDAC5 (Cell Signaling Technology, 2082), HDAC6 (Santa Cruz Biotechnology, 11420), HDAC7 (Cell Signaling Technology, 2882), calnexin (Santa Cruz Biotechnology, 11397).
Blakeslee W.W., Demos-Davies K.M., Lemon D.D., Lutter K.M., Cavasin M.A., Payne S., Nunley K., Long C.S., McKinsey T.A, & Miyamoto S.D. (2017). Histone Deacetylase Adaptation in Single Ventricle Heart Disease and a Young Animal Model of Right Ventricular Hypertrophy. Pediatric research, 82(4), 642-649.
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2

Western Blot Analysis of Cellular Proteins

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For protein analysis, 5–30 μg of whole cell protein extract (WCE) were loaded into 6%–15% 37.5:1 bis-acrylamide SDS-PAGE gels, depending on protein of interest size. Antibodies for Western blot were diluted in 5% BSA-TBST as follows: HDAC7 (Cell Signaling D4E1L, 1:1000), Lamin-B1 (Abcam ab133741; 1:5000), PKM2 (Cell Signaling D78A4; 1:1000), Flag (Cell Signaling, 1:1000-2000), pan histone H3 (Abcam ab1791), H3K9ac (Abcam ab4441), H3K27ac (Abcam ab4729), HNRNPL (4D11, Abcam ab6106).
Agosto L.M., Mallory M.J., Ferretti M.B., Blake D., Krick K.S., Gazzara M.R., Garcia B.A, & Lynch K.W. (2023). Alternative splicing of HDAC7 regulates its interaction with 14-3-3 proteins to alter histone marks and target gene expression. Cell reports, 42(3), 112273.
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3

Epigenetic Regulation of Nrf2 Pathway

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CRA (≥ 98% in purity) was purchased from Chendu Biopurify Medical Technology Co., Ltd. (Chendu, China; 13120404). 5-Aza-2′-deoxycytidine (5-AZA), trichostatin A (TSA), ampicillin, bovine serum albumin (BSA), and a protein inhibitor cocktail were purchased from Sigma-Aldrich (St. Louis, MO, USA). Dulbecco’s modified Eagle’s medium (DMEM), fetal bovine serum (FBS), and trypsin-EDTA (0.25%) were purchased from Gibco (Carlsbad, CA, USA). The anti-Nrf2, anti-HO-1, and anti-NQO-1 antibodies were purchased from Abcam (Cambridge, MA, USA). Anti-DNMT1, anti-DNMT3a and anti-DNMT3b antibodies were supplied by Novus Biologicals (Littleton, CO, USA). The antibodies against HDACs (HDAC1, HDAC2, HDAC4, HDAC6 and HDAC7) were purchased from Cell Signaling Technology (Boston, MA, USA). The anti-HDAC8 antibody was obtained from Proteintech Group (Chicago, IL, USA). The anti-β-actin primary antibody and secondary antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).
Yang J., Wu R., Li W., Gao L., Yang Y., Li P, & Kong A.N. (2018). The triterpenoid corosolic acid blocks transformation and epigenetically reactivates Nrf2 in TRAMP-C1 prostate cells. Molecular carcinogenesis, 57(4), 512-521.
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4

Evaluating HDAC Protein Expression

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Cells were harvested using lysis buffer (iNtRON Biotechnology, Seoul, Korea). Proteins were solubilized by sonication and equal amounts of protein were separated on sodium dodecyl sulphate polyacrylamide gel electrophoresis and electroblotted onto polyvinylidene difluoride membranes (Millipore, Bedford, MA). Membranes were blocked in phosphate-buffered saline containing 0.1% Tween 20 and 5% powdered milk, and probed with the primary antibody against each HDAC isotype (HDAC1, HDAC3, HDAC4, HDAC5, HDAC6, and HDAC7 antibodies from Cell Signaling, Beverly, MA; HDAC8 and HDAC9 antibodies from Santa Cruz Biotechnology, Santa Cruz, CA; HDAC10 antibody from Sigma-Aldrich, St. Louis, MO; HDAC11 antibody from Abgent, San Diego, CA). Antibodies against Rad51, phospho-ataxia telangiectasia mutated (phospho-ATM), Aurora A, Aurora B, X-linked inhibitor of apoptosis (XIAP), and survivin were purchased from Cell Signaling Technology.
Kim J.H., Moon S.H., No M., Kim J.J., Choi E.J., Cho B.J., Kim J.S., Kim I.H, & Kim I.A. (2015). Isotype-Specific Inhibition of Histone Deacetylases: Identification of Optimal Targets for Radiosensitization. Cancer Research and Treatment : Official Journal of Korean Cancer Association, 48(3), 1130-1140.
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5

Generation and Validation of Phospho-Specific Antibodies

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Rabbit polyclonal phospho-specific antibodies against MARK2 S456, S569, S619, and MST2 S15 were generated and purified by AbMart, Inc. The peptides used for immunizing rabbits were AKVPA-pS-PLPGL (MAR2 S456), RVPVA-pS-PSAHN (MARK2 S569), GVTPA-pS-PSGHS (MARK2 S619), and KLKKL-pS-EDSLT (MST2 S15). The corresponding non-phosphorylated peptides were also used for antibody purification and blocking assays. Anti-MARK2, Cyclin B1, β-actin were purchased from Santa Cruz Biotechnology. Anti-MARK1, MARK3, MARK4, p-MARK T208 (activation loop), HDAC1, HDAC2, HDAC4, HDAC5, HDAC6, HDAC7, p-HDAC4/5/7(S246/S259/S155), p-HDAC4/5/7(S632/S661/S486), cleaved-PARP (human specific), cleaved-PARP (rodent specific), cleaved-caspase 3, Erk1/2, Zyxin, Survivin, p-YAP S127, p-YAP S397, YAP, and p-Aurora-A/B/C were from Cell Signaling Technology. Anti-GST and LATS2 antibodies were from Bethyl Laboratory. Anti-Aurora-A and Flag antibodies were from Sigma. Anti-α-tubulin antibody was from Abcam Inc. All other antibodies used in this study are listed in Table S1.
Zeng Y., Yin L., Zhou J., Zeng R., Xiao Y., Black A.R., Hu T., Singh P.K., Yin F., Batra S.K., Yu F., Chen Y, & Dong J. (2022). MARK2 Regulates Chemotherapeutic Responses through Class IIa HDAC-YAP Axis in Pancreatic Cancer. Oncogene, 41(31), 3859-3875.
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6

Immunoblotting of HDAC Isoforms

Cited 1 time
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Immunoblotting was performed as previously described(16 (link)). Briefly, pediatric human and neonatal rat RV homogenates were prepared and concentrations quantified as above for HDAC catalytic activity assay. Proteins were resolved by SDS-PAGE, transferred to nitrocellulose membranes (BioRad) and probed with antibodies for HDAC1 (Cell Signaling Technology, 5356), HDAC2 (Cell Signaling Technology, 5113), HDAC3 (Cell Signaling Technology, 3949), HDAC4 (Cell Signaling Technology, 5392), HDAC5 (Cell Signaling Technology, 2082), HDAC6 (Santa Cruz Biotechnology, 11420), HDAC7 (Cell Signaling Technology, 2882), calnexin (Santa Cruz Biotechnology, 11397).
Blakeslee W.W., Demos-Davies K.M., Lemon D.D., Lutter K.M., Cavasin M.A., Payne S., Nunley K., Long C.S., McKinsey T.A, & Miyamoto S.D. (2017). Histone Deacetylase Adaptation in Single Ventricle Heart Disease and a Young Animal Model of Right Ventricular Hypertrophy. Pediatric research, 82(4), 642-649.
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7

Proteasome Inhibition and mTOR Regulation

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N-(2,5-Dichlorophenyl)-4-(furan-2-yl)-2-methyl-5-oxo-1,4,5,6,7,8-hexahydro-quinoline-3-carboxamide (AR420626) and MG-132 (Sigma-Aldrich, St. Louis, MO, USA); TNF-α antagonist R-705013 (link) (Calbiochem, Darmstadt, Germany); rapamycin (Selleckchem, Houston, TX, USA); polyclonal rabbit antibodies against human β-actin (Abcam, Cambridge, UK), acetyl-histone H3 (Lys9/Lys14), cleaved caspase-3 (Asp175), cleaved caspase-9, phospho-mTOR (Ser2448) (Cell Signaling Technology, Boston, MA, USA); monoclonal rabbit antibodies against HDAC1, HDAC2, HDAC4, HDAC5, HDAC6, HDAC7, mTOR (Cell Signaling Technology) and HDAC8 (Abcam); monoclonal mouse antibodies against HDAC3, caspase-8 (Cell Signaling Technology) and 20S Proteasome β5 (Santa Cruz Biotechnology, Dallas, TX, USA); and horseradish peroxidase (HRP)-conjugated antimouse and antirabbit immunoglobulins (Dako, Glostrup, Denmark) were used in the study.
Mikami D., Kobayashi M., Uwada J., Yazawa T., Kamiyama K., Nishimori K., Nishikawa Y., Nishikawa S., Yokoi S., Taniguchi T, & Iwano M. (2020). AR420626, a selective agonist of GPR41/FFA3, suppresses growth of hepatocellular carcinoma cells by inducing apoptosis via HDAC inhibition. Therapeutic Advances in Medical Oncology, 12, 1758835920913432.
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8

Western Blot Analysis of Cellular Signaling

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Cells in spheroids were lysed and the whole cell lysates were determined by10% sodium dodecyl sulphate polyacrylamide gel electrophoresis and polyvinylidene difluoride (PVDF) membranes transfer. The membranes were incubated with specific primary antibodies at 4°C overnight and HRP‐conjugated secondary antibodies at room temperature for 2 h. The primary antibodies were including rabbit anti‐HDAC6 (#7558), HDAC7 (#10831), total STAT3 (#12640), total ERK (#4695), total JNK (#9258), phospho‐STAT3 (#9145), phosphor‐ERK (#4370), phospho‐JNK (#4668), COX2 (#12282) and GAPDH (Cell Signalling Technology, Beverly, MA, USA).
Ou L., Wang H., Huang H., Zhou Z., Lin Q., Guo Y., Mitchell T., Huang A.C., Karakousis G., Schuchter L., Amaravadi R., Guo W., Salvino J., Herlyn M, & Xu X. (2022). Preclinical platforms to study therapeutic efficacy of human γδ T cells. Clinical and Translational Medicine, 12(6), e814.
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9

Western Blot Analysis of Cellular Proteins

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For protein analysis, 5–30 μg of whole cell protein extract (WCE) were loaded into 6%–15% 37.5:1 bis-acrylamide SDS-PAGE gels, depending on protein of interest size. Antibodies for Western blot were diluted in 5% BSA-TBST as follows: HDAC7 (Cell Signaling D4E1L, 1:1000), Lamin-B1 (Abcam ab133741; 1:5000), PKM2 (Cell Signaling D78A4; 1:1000), Flag (Cell Signaling, 1:1000-2000), pan histone H3 (Abcam ab1791), H3K9ac (Abcam ab4441), H3K27ac (Abcam ab4729), HNRNPL (4D11, Abcam ab6106).
Agosto L.M., Mallory M.J., Ferretti M.B., Blake D., Krick K.S., Gazzara M.R., Garcia B.A, & Lynch K.W. (2023). Alternative splicing of HDAC7 regulates its interaction with 14-3-3 proteins to alter histone marks and target gene expression. Cell reports, 42(3), 112273.
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10

Epigenetic Modulators and Cellular Signaling

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TAX (purity ≥ 98%), 5-azadeoxycytidine (5-aza, purity ≥ 97%), trichostatin A (TSA, purity ≥ 98%), bacteriological agar, Eagle’s basal medium (BME) and TPA were obtained from Sigma-Aldrich (St. Louis, MO, USA). Fetal bovine serum (FBS), Dulbecco’s modification of eagle’s medium (DMEM), minimum essential medium (MEM) and trypsin-EDTA solution were obtained from Gibco Laboratories (Gaithersburg, MD, USA). The primary antibodies anti-Nrf2, anti-HO-1, anti-NQO-1 and anti-β-actin were acquired from Santa Cruz Biotechnology (Santa Cruz, CA, USA). The primary antibodies anti-DNMT (DNMT1, DNMT3a and DNMT3b) were obtained from IMGENEX (San Diego, CA, USA). The primary antibodies anti-HDAC (HDAC1, HDAC2, HDAC3, HDAC4, HDAC5, HDAC6, HDAC7 and HDAC8) were obtained from Cell Signaling Technology (Danvers, MA, USA).
Kuang H., Tang Z., Zhang C., Wang Z., Li W., Yang C., Wang Q., Yang B, & Kong A.N. (2017). Taxifolin Activates the Nrf2 Anti-Oxidative Stress Pathway in Mouse Skin Epidermal JB6 P+ Cells through Epigenetic Modifications. International Journal of Molecular Sciences, 18(7), 1546.
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