ChIPAb+ HDAC3 Antibody

49-53kDa

FUNCTION: SwissProt: O15379 # Responsible for the deacetylation of lysine residues on the N-terminal part of the core histones (H2A, H2B, H3 and H4). Histone deacetylation gives a tag for epigenetic repression and plays an important role in transcriptional regulation, cell cycle progression and developmental events. Histone deacetylases act via the formation of large multiprotein complexes. Probably participates in the regulation of transcription through its binding to the zinc-finger transcription factor YY1; increases YY1 repression activity. Required to repress transcription of the POU1F1 transcription factor.
SIZE: 428 amino acids; 48848 Da
SUBUNIT: Interacts with HDAC7 and HDAC9. Forms a heterologous complex at least with YY1. Interacts with DAXX, HDAC10 and DACH1. Found in a complex with NCOR1 and NCOR2. Component of the N-Cor repressor complex, at least composed of NCOR1, NCOR2, HDAC3, TBL1X, TBL1R, CORO2A and GPS2. Interacts with BCOR, MJD2A/JHDM3A, NRIP1 and SRY. Interacts with BTBD14B. Interacts with GLIS2 (By similarity). Interacts with CBFA2T3.
SUBCELLULAR LOCATION: Nucleus.
TISSUE SPECIFICITY: Widely expressed.
PTM: Sumoylated in vitro.
SIMILARITY: SwissProt: O15379 ## Belongs to the histone deacetylase family. Type 1 subfamily.

peptide (NEFYDGDHDNDKESDVEI) corresponding to amino acids 411-428 of human HDAC3

Chromatin Immunoprecipitation:

Representative lot data.

Sonicated chromatin prepared from untreated or UV treated (6 hrs, 50 joules/m2.) U2OS cells (3 X 10E6 cell equivalents per IP) were subjected to chromatin immunoprecipitation using 4 µg of either Normal Mouse IgG, or 4 µg of Anti-HDAC3 and the MAGNA CHIP^® A/G Kit (Cat. # 17-10085). Successful immunoprecipitation of HDAC3 associated DNA fragments was verified by qPCR using ChIP Primers, p21 .(Figure 2). Data is presented as percent input of each IP sample relative to input chromatin for each amplicon and ChIP sample as indicated.

Please refer to the EZ-MAGNA CHIP^® A (Cat. # 17-408) or EZ-ChIP (Cat. # 17-371) protocol for experimental details.

Western Blot Analysis:

Representative lot data.

HeLa nuclear extract was probed with Anti-HDAC3 (1 µg/mL). Proteins were visualized using a goat anti-mouse secondary antibody conjugated to HRP and a chemiluminescence detection system.

Arrow indicates HDAC3 (~49 kDa). An uncharacterized band appears at ~22 in some lysates. (Figure 3).

25 assays per set. Recommended use: 4 μg of antibody per chromatin immunoprecipitation (dependent upon biological context).

Anti-HDAC3 (mouse monoclonal). One vial containing 100 µg of protein G purified monoclonal in buffer containing 0.02 M phosphate buffer (pH7.6), 0.25M NaCl, 0.1% sodium azide and 30% glycerol. Store at -20°C.
Concentration: 1 mg/mL
Normal Mouse IgG. One vial containing 125 µg of purified mouse IgG in 125 µL of storage buffer containing 0.1% sodium azide. Store at -20°C.
ChIP Primers, p21. One vial containing 75 μL of each primer (5 μM) specific for a region of the human p21 (WAF1/CIP1/CDKN1A) promoter. Store at
-20°C.
FOR: CCC ACA GCA GAG GAG AAA GAA
REV: CTG GAA ATC TCT GCC CAG ACA

Stable for 1 year at -20°C from date of receipt. Handling Recommendations: Upon first thaw, and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance. Note: Variability in freezer temperatures below -20°C may cause glycerol containing solutions to become frozen during storage.

Demonstrated to react with human, bovine, human, and hamster. Predicted to react with rat and chicken based on 100% sequence homology.

Includes normal mouse IgG and primers specific for human p21.

MAGNA CHIP is a registered trademark of Merck KGaA, Darmstadt, Germany

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