Hdac1

Manufactured by BPS Biosciences

Sourced in United States

HDAC1 is a recombinant human histone deacetylase 1 enzyme. Histone deacetylases are a class of enzymes that remove acetyl groups from lysine residues on histone proteins, which can lead to changes in chromatin structure and gene expression.

Automatically generated - may contain errors

Lab products found in correlation

Hdac6, by BPS Biosciences (4 mentions) Hdac2, by BPS Biosciences (3 mentions) Hdac4, by BPS Biosciences (2 mentions) Hdac3, by BPS Biosciences (2 mentions) Hdac assay developer, by BPS Biosciences (1 mentions) Bovine serum albumin (bsa), by Merck Group (1 mentions) Hrp conjugated anti rabbit antibody, by Cell Signaling Technology (1 mentions) Anti h3k27ac, by Cell Signaling Technology (1 mentions) Anti h4k16ac, by Active Motif (1 mentions) Anti histone h3, by Abcam (1 mentions) Anti h3k9ac, by Cell Signaling Technology (1 mentions) His tag monoclonal antibody, by Thermo Fisher Scientific (1 mentions) Dimethyl sulfoxide (dmso), by Merck Group (1 mentions) Penicillin, by Thermo Fisher Scientific (1 mentions) Streptomycin, by Thermo Fisher Scientific (1 mentions) Hdac6, by Cell Signaling Technology (1 mentions) Glyceraldehyde 3 phosphate dehydrogenase, by Cell Signaling Technology (1 mentions) Hdac11, by BPS Biosciences (1 mentions) Hdac fluorogenic assay kits, by BPS Biosciences (1 mentions) Amersham ecl select western blotting detection reagent, by GE Healthcare (1 mentions)

Close spelling variants of this product

HDAC1 (6xHis/FLAG; Recombinant human HDAC1 Recombinant HDAC1

8 protocols using hdac1

HDAC Enzyme Inhibition Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols

The HDAC assay kit was purchased from BPS Bioscience (San Diego, CA). The assays were carried out in black NUNC 96-well plates. Each well contained a volume of 50 μl including buffer (BPS Bioscience, catalogue no. 50031), the mixture of proteins (HDAC-1 (0.033 mg/ml, BPS Bioscience, catalogue no. 50051) or HDAC-6 (0.025 mg/ml, BPS Bioscience, catalogue no. 50006) and competing proteins), inhibitor (at the IC50), BSA (1 mg/mL, Sigma-Aldrich), and HDAC substrate 3 (20 μM, BPS Bioscience, catalogue no. 50037). Upon addition of substrate, the plate was incubated at 37 °C for 30 min. HDAC assay developer (50 μL, BPS Bioscience, catalogue no. 50030) was then added to each well and the plate incubated for 15 min at room temperature. The fluorescence was recorded at excitation and emission wavelengths of 360 and 460 nm, respectively. Blank wells containing no inhibitor or protein were subtracted from all wells. The control wells, containing no inhibitor, were arbitrarily set as 100% activity. The assays were performed in triplicate, and each assay contained each inhibitor/competing protein combination was conducted three times. The data were normalized to values measured for uninhibited enzyme. Assays were reported as mean ± standard deviation.

Chen Y, & Cohen S.M. (2015). Investigating the Selectivity of Metalloenzyme Inhibitors in the Presence of Competing Metalloproteins. ChemMedChem, 10(10), 1733-1738.

+ Open protocol

+ Expand

Histone Acetylation Profiling

Check if the same lab product or an alternative is used in the 5 most similar protocols

20 amino acids were purchased from EMP Millipore. Site specific primary antibodies include anti-H3K9ac (Cell Signaling), anti-H3K14ac (EMP Millipore), anti-H3K18ac (EMP Millipore), anti-H3K23ac (EMP Millipore), anti-H3K27ac (Cell Signaling), anti-H4K16ac (Active Motif) antibodies, as well as anti-histone H3 (Abcam). Secondary antibody as HRP-conjugated anti-rabbit antibody from Cell Signaling. H4K16ac nucleosome was purchased from Epicypher. The free enzyme HDAC1 (in the complex with HSP70) is purchased from BPS Bioscience.

Wang Z.A., Millard C.J., Lin C.L., Gurnett J.E., Wu M., Lee K., Fairall L., Schwabe J.W, & Cole P.A. (2020). Diverse nucleosome Site-Selectivity among histone deacetylase complexes. eLife, 9, e57663.

+ Open protocol

+ Expand

HDAC Inhibition Assay Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols

HDAC-1 and −6 were purchased from BPS Bioscience (BPS Bioscience catalog #50051 and 50006, San Diego, CA, USA) and the assay was carried out as instructed by manufacturer. The assays were carried out in black 96-well plates (Costar). Each well contained a volume of 50 μL including buffer (25 mM Tris/HCl, 137 mM NaCl, 2.7 mM KCl, 1 mM MgCl₂, 0.1 mg/mL BSA, pH 8.0), HDAC (3.8 ng/well of HDAC-1, 50 ng/well of HDAC-6, BPS Bioscience, catalogue no. 50051 and no. 50006, respectively), inhibitor (various concentrations, 1 mM – 5 μM), and HDAC substrate 3 (20 μM, BPS Bioscience, catalogue no. 50037). Prior to adding substrate, the plate was preincubated for 5 min. Upon addition of substrate, the plate was incubated at 37 °C for 30 min. HDAC assay developer (50 μL, BPS Bioscience, catalogue no. 50030) was added to each well and the plate incubated for 15 min at room temperature. The fluorescence was recorded at excitation and emission wavelengths of 360 and 460 nm, respectively. The negative control wells, containing no inhibitor, were arbitrarily set as 100% activity. The positive control wells, containing 200 μM SAHA, were arbitrarily set as 0% activity. HDAC activity was defined as the ratio of fluorescence in the inhibitor wells relative to the negative control wells, expressed as a percentage. The assays were performed in triplicate.

Chen A.Y., Thomas P.W., Stewart A.C., Bergstrom A., Cheng Z., Miller C., Bethel C.R., Marshall S.H., Credille C.V., Riley C.L., Page R.C., Bonomo R.A., Crowder M.W., Tierney D.L., Fast W, & Cohen S.M. (2017). Dipicolinic Acid Derivatives as Inhibitors of New Delhi Metallo-β-lactamase-1. Journal of medicinal chemistry, 60(17), 7267-7283.

+ Open protocol

+ Expand

HDAC1 Inhibition Assay Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols

The enzymatic test against HDAC1 was performed by Chempartner (Shanghai, China). The compounds were tested in vitro using SAHA (purchased from Sigma -Aldrich, Merck KGaA, Darmstadt, Germany and/or its affiliates, Cat. No. SML0061) as the reference compound, and the testing concentration was set as 100 μM. HDAC1 was purchased from BPS bioscience (San Diego, CA, USA) and prepared in modified Tris Buffer. The substrate solution was made by trypsin and Ac-peptide substrate in modified Tris Buffer. A total of 15 μL of the HDAC1 solution was transferred to the assay plate and incubated at room temperature for 15 min; then, 10 μL of the substrate solution was added to each well to start the reaction. The fluorescence was then measured for excitation and emission at wavelengths of 355 nm and 460 nm by a Envision plate reader. The percentage inhibition was calculated using the following equation: Inhibition % = (Max-Signal)/(Max-Min) × 100%. All the tests were performed in duplicate.

Zhang L., Yang Y., Yang Y, & Xiao Z. (2024). Discovery of Novel Metalloenzyme Inhibitors Based on Property Characterization: Strategy and Application for HDAC1 Inhibitors. Molecules, 29(5), 1096.

+ Open protocol

+ Expand

HDAC Enzyme Characterization Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols

Assays were performed in 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES) buffer [50 mM HEPES/Na, 100 mM KCl, 0.001% (v/v) tween-20, 0.2 mM tris(2-carboxyethyl)phosphine (TCEP), 0.5 mg/mL bovine serum albumin (BSA), pH 7.4] unless otherwise stated. Recombinant HDAC enzymes employed in µSPOT, inhibitor, and substrate assays were from commercial sources: HDAC1 (full length, C-terminal His tag, C-terminal FLAG tag, BPS Bioscience, cat. #: 50051, lots 170105-1 and 181108-1, purity ≥ 79%, activity ≥ 460 pmol/min/µg), HDAC2 (full length, C-terminal His tag, BPS Bioscience, cat. #: 50002, lots 160701 and 160630, purity ≥ 84%, activity ≥ 675 pmol/min/µg), HDAC3/NCoR2 (full length, C-terminal His tag, NCoR2 N-terminal GST tag, BPS Bioscience, cat. #: 50003, lots 130819 and 190327, purity ≥ 80%, activity ≥ 3000 pmol/min/µg), HDAC8 (full length, C-terminal His tag, BPS Bioscience, cat. #: 50008, lots 150714 and 161216, purity ≥ 90%, activity ≥ 300 pmol/min/µg), HDAC4 (aa 627‒1084, BPS Bioscience, cat. #: 50004, lot 130828-G, purity ≥ 89%, activity ≥ 103225 pmol/min/µg), HDAC6 (full length, BPS Bioscience, cat. #: 50056, lot 151130-C, purity ≥ 88%, activity ≥ 150 pmol/min/µg). Antibodies: 6x-His tag monoclonal antibody (HIS.H8), HRP-conjugated (ThermoFisher, cat. #: MA1-21315-HRP).

Moreno-Yruela C., Bæk M., Vrsanova A.E., Schulte C., Maric H.M, & Olsen C.A. (2021). Hydroxamic acid-modified peptide microarrays for profiling isozyme-selective interactions and inhibition of histone deacetylases. Nature Communications, 12, 62.

+ Open protocol

+ Expand

HDAC1 Enzyme Activity Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols

HDAC1 (BPS Bioscience Inc., San Diego, CA, USA. Cat. No. 50051); DMSO (Sigma, St. Louis, MO, USA. Cat. No. D2650); 84-well plate (Perkin Elmer Inc., Waltham, MA, USA. Cat. No. 6007279)

Lin Y., Zhang H., Niu T., Tang M.L, & Chang J. (2020). Discovery of Novel Indoleamine 2,3-Dioxygenase 1 (IDO1) and Histone Deacetylase 1 (HDAC1) Dual Inhibitors Derived from the Natural Product Saprorthoquinone. Molecules, 25(19), 4494.

+ Open protocol

+ Expand

Histone Deacetylase Assays and Immunoblotting

Check if the same lab product or an alternative is used in the 5 most similar protocols

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.

+ Open protocol

+ Expand

HDAC Enzyme Activity Assay Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols

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.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!