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Hdac4

Manufactured by BPS Biosciences
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HDAC4 is a recombinant human histone deacetylase enzyme. Histone deacetylases are enzymes that remove acetyl groups from lysine residues on histone proteins, leading to changes in chromatin structure and gene expression.

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Lab products found in correlation

Hdac2, by BPS Biosciences (3 mentions) Hdac6, by BPS Biosciences (3 mentions) Hdac3, by BPS Biosciences (3 mentions) Hdac9, by BPS Biosciences (2 mentions) Hdac5, by BPS Biosciences (2 mentions) Hdac7, by BPS Biosciences (2 mentions) Hdac8, by BPS Biosciences (2 mentions) Hdac1, by BPS Biosciences (2 mentions) Fluorogenic hdac assay kit, by BPS Biosciences (1 mentions) Spectramax gemini, by Molecular Devices (1 mentions) Epidermal growth factor (egf), by Thermo Fisher Scientific (1 mentions) Celltiter glo reagent, by Promega (1 mentions) Fibronectin, by Thermo Fisher Scientific (1 mentions) Stempro neural supplement, by Thermo Fisher Scientific (1 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions) Trypsin edta, by Thermo Fisher Scientific (1 mentions) Hepg2, by American Type Culture Collection (1 mentions) Hct116, by American Type Culture Collection (1 mentions) Hdac activity assay kit, by Cayman Chemical (1 mentions) Hdac1, by Cayman Chemical (1 mentions)

5 protocols using hdac4

1

Fluorogenic HDAC Enzyme Assay

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Enzyme-active recombinant HDAC4, 7 and 8 were purchased from BPS Bioscience Inc., and HDAC2 was purchased from BIOMOL. The HDAC4 and HDAC7 assays were performed by Fluorogenic HDAC Class 2a Assay Kit (BPS Bioscience), and the HDAC2 and HDAC8 assays were performed by Fluorogenic HDAC Assay Kit (BPS Bioscience). The specific HDAC fluorometric substrate, comprised of an acetylated lysine side chain, was incubated with purified individual HDAC enzymes (HDAC2, 4, 7 and 8), in the presence or absence of the HDAC inhibitors, PBA, BS, TSA or VAL, at different doses. After adding Lysine Developer, the fluorophore that could then be measured using the fluorescence plate reader Spectra (Max Gemini, Molecular Devices, Sunnyvale, CA), was excitated at a wave-length of 365 nm and detected by emitted light at 450 nm.
Ma J., Luo T., Zeng Z., Fu H., Asano Y., Liao Y., Minamino T, & Kitakaze M. (2016). Histone Deacetylase Inhibitor Phenylbutyrate Exaggerates Heart Failure in Pressure Overloaded Mice independently of HDAC inhibition. Scientific Reports, 6, 34036.
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2

High-Throughput Screening of Epigenetic Modulators

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Four cell lines (human colon cancer cell line HCT116, human embryonic kidney cell line HEK293, human liver cancer cell line HepG2, and human B lymphoma cell line SU-DHL-6), fetal bovine serum (FBS), Eagle's Minimum Essential Medium (EMEM), and Roswell Park Memorial Institute (RPMI) 1640 medium were purchased from American Type Cell Collection (ATCC, Manassas, VA, USA). H9-derived human neural stem cells, defined FBS, trypsin-EDTA, fibronectin, basal fibroblast growth factor (bFGF), epidermal growth factor (EGF), KnockOut™ Dulbecco's Modified Eagle Medium (DMEM)/F12, and penicillin-streptomycin, StemPro® Neural Supplement were acquired from Life Technologies, Carlsbad, CA, USA. Chemicals and epigenetic compound libraries were purchased from Cayman Chemicals (Ann Arbor, MI), Santa Cruz Biotechnology (Dallas, TX, USA), SelleckChem (Houston, TX, USA), and Sigma-Aldrich (St. Louise, MO, USA). HDAC-Glo I/II and CellTiter-Glo reagents were purchased from Promega, Madison, WI. Fluorogenic assay kits for HDAC1, HDAC2, HDAC3, HDAC4, HDAC5, HDAC6, HDAC7, HDAC8, HDAC9, and HDAC10 were acquired from BPS Bioscience (San Diego, CA). White solid bottom 1536-well assay plates were purchased from Greiner Bio-One (Monroe, NC).
Hsu C.W., Shou D., Huang R., Khuc T., Dai S., Zheng W., Klumpp-Thomas C, & Xia M. (2016). Identification of HDAC inhibitors using a cell-based HDAC I/II assay. Journal of biomolecular screening, 21(6), 643-652.
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3

Measuring HDAC Activity Assay

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HDAC activity of class I HDACs (HDAC1, 2, 3 and 8) was measured with the fluorometric HDAC Activity Assay kit: HDAC1 (10011563, Cayman Chemical, Ann Arbor, MI), HDAC2, HDAC 3, and HDAC 8 (50062, 50073 and 50068, BPS Bioscience, San Diego, CA), according to the manufacturer’s instructions. HDAC activity of class II HDACs (HDAC4 and HDAC6) were measured with the fluorometric HDAC Activity Assay kit: HDAC4, HDAC 4 (50064 and 50076, BPS Bioscience, San Diego, CA), according to the manufacturer’s instructions.
Due to interference, concentrations of >30 mM were not used in the IC50 calculation. To account for any interference at lower concentrations, all baseline measurements of blank wells (no HDAC enzyme) were performed and subtracted with the various concentrations of diacetyl.
Haga-Yamanaka S., Nunez-Flores R., Scott C.A., Perry S., Chen S.T., Pontrello C., Nair M.G, & Ray A. (2023). Plasticity of gene expression in the nervous system by exposure to environmental odorants that inhibit HDACs. bioRxiv.
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4

Histone Deacetylase Assays and Immunoblotting

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Dulbecco's modified Eagle's medium (DMEM) with L-glutamine was purchased from GenDEPOT (Barker, TX), fetal bovine serum was purchased from HyClone, and penicillin and streptomycin were purchased from Gibco BRL (Gaithersburg, MD). Antibodies for α-tubulin, Ac-α-tubulin (Lys40), Histone H3, Ac-Histone H3 (Lys9), HDAC1, HDAC6, and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) were purchased from Cell Signaling Technology (Boston, MA). Goat anti-rabbit IgG horseradish peroxidase conjugate was purchased from Santa Cruz Biotechnology (Santa Cruz, CA). Cell Titer 96 Aqueous One Solution cell proliferation assay kit was purchased from Promega (Madison, WI). Amersham ECL Select western blotting Detection Reagent was purchased from GE Healthcare. HDAC fluorogenic assay kits (HDAC1, #50061; HDAC2, #50062; HDAC3, #50073; HDAC4, #50064; HDAC6, #50076; HDAC11, #50687) were bought from BPS Bioscience (San Diego, CA).
Kim J.H., Ali K.H., Oh Y.J, & Seo Y.H. (2022). Design, synthesis, and biological evaluation of histone deacetylase inhibitor with novel salicylamide zinc binding group. Medicine, 101(17), e29049.
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5

HDAC Enzyme Activity Assay Protocol

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Enzyme activity measurements of class I and IIa HDACs were performed using a cell‐free system and according to a method previously described.16 We used fluorogenic HDAC1 (#50061), HDAC2 (#50062), HDAC3 (#50063), HDAC4 (#50064), HDAC5 (#50065), HDAC6 (#50076), HDAC7 (#50067), HDAC8 (#50068) and HDAC9 (#50069) enzyme assay kits from BPS Bioscience (San Diego, CA, USA). To test the enzyme activity of LMK235, 0.001, 0.003, 0.01, 0.03, 0.1, 0.3, 1, 3 and 10 μmol/L concentrations were used. For the IC50 calculations, every data point was normalized to the vehicle (100% activity). The normalized data were fitted with a Hill non‐linear curve fit (OrigionPro 9.0). Employing the “Find X from Y” function in OrigionPro 9.0 resulted in the IC50 values (50% activity).
Choi S.Y., Kee H.J., Sun S., Seok Y.M., Ryu Y., Kim G.R., Kee S., Pflieger M., Kurz T., Kassack M.U, & Jeong M.H. (2019). Histone deacetylase inhibitor LMK235 attenuates vascular constriction and aortic remodelling in hypertension. Journal of Cellular and Molecular Medicine, 23(4), 2801-2812.
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