AbFlex® HDAC2 antibody (rAb)

RRID: AB_2793799
Catalog No: 91197 Format: 100 µg $350 Buy
Catalog No: 91198 Format: 10 µg $75 Buy

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Antibody Type:
Recombinant
Isotype:
IgG2a
Purification:
Protein A Chromatography
Host:
Mouse
Molecular Weight:
55 kDa
Reactivity:
Human

Applications

ChIP Validated ChIP-Seq Validated Western Blot Validated

Application Notes

Applications Validated by Active Motif:
ChIP-Seq: 4 - 10 µg per ChIP
WB*: 0.2-2 µg/ml

For optimal results, primary antibody incubations should be performed at room temperature. The addition of 0.1% Tween 20 to all blocking solutions may also reduce background. Individual optimization may be required.

*Note: many chromatin-bound proteins are not soluble in a low salt nuclear extract and fractionate to the pellet. Therefore, we recommend a High Salt / Sonication Protocol when preparing nuclear extracts for Western Blot.

Immunogen

This antibody was raised against peptide corresponding to amino acids 473-488 of human HDAC2.

Buffer

Purified IgG in 140 mM Hepes, pH 7.5, 70 mM NaCl, 35 mM NaOAc, 0.07% sodium azide, 30% glycerol.

AbFlex® recombinant antibodies are genetically derived from DNA sequences of parental hybridoma clones.  For details on the parental clone, see Catalog No. 39533.

 

AbFlex® HDAC2 recombinant antibody (rAb) tested by ChIP-Seq
Chromatin immunoprecipitation (ChIP) was performed using the ChIP-IT® High Sensitivity Kit (Cat. No. 53040) with 30 µg of chromatin from NCI-H1437 (human lung carcinoma cell line) chromatin and 4 µg HDAC2 antibody. ChIP DNA was sequenced on the Illumina HiSeq and 15.6 million sequence tags were mapped to identify HDAC2 binding sites on chromosome 1. You can view the complete data set in the UCSC Genome Browser, starting at this specific location, here.

Western blot of AbFlex® HDAC2 antibody (rAb).
20 µg HeLa cell nuclear extract was run on SDS-PAGE and probed with 0.5 µg/ml AbFlex® HDAC2 antibody (rAb).

MW: 55 kDa

Background

AbFlex® antibodies are recombinant antibodies (rAbs) that have been generated using defined DNA sequences to produce highly specific, reproducible antibodies. Each AbFlex antibody contains a 6xHis Tag, a Biotinylation Tag for enzymatic biotin conjugation using the biotin ligase, BirA, and a sortase recognition motif (LPXTG) to attach a variety of labels directly to the antibody including fluorophores, enzymatic substrates (HRP, AP), peptides, drugs as well as solid supports.

HDAC2 (Histone Deacetylase 2, also designated mammalian RPD3) is a member of the class I mammalian histone deacetylases (HDACs) involved in regulating chromatin structure during transcription. These enzymes catalyze the removal of acetyl groups from lysine residues of histones and other cellular proteins. Lysine N-ε-acetylation is a dynamic, reversible and tightly regulated protein and histone modification that plays a major role in regulation of gene expression in various cellular functions. It consists of the transfer of an acetyl moiety from an acetyl coenzyme A to the ε-amino group of a lysine residue.

In vivo, acetylation is controlled by the antagonistic activities of histone acetyltransferases (HATs) and histone deacetylases (HDACs). The HDACs are grouped into four classes, on the basis of similarity to yeast counterparts: HDAC class I (HDAC1, HDAC2, HDAC3 and HDAC8), class II (HDAC4, HDAC5, HDAC6, HDAC7, HDAC9 and HDAC10), class III (SIRT1, SIRT2, SIRT3, SIRT4, SIRT5, SIRT6 and SIRT-7) and class IV (HDAC11).

HDAC2 is associated with many different proteins as YY1 (a mammalian zinc-finger transcription factor). HDAC2 also forms transcriptional repressor complexes containing, among others, HDAC1 or RBBP4. HDAC1, HDAC2 and HDAC3 are also ubiquitously expressed and can deacetylate both Histone H3 and Histone H4 in free histones or nucleosome substrate.

Chromatin IP Assays

This antibody has been validated for use in ChIP and/or ChIP-Seq, and can be used with Active Motif's ChIP-IT® High Sensitivity Kit or our magnetic bead-based ChIP-IT® Express Kits. For an overview of all of our ChIP products, please visit our Chromatin IP Page to learn about ChIP products designed for use with AbFlex® HDAC2 antibody (rAb).

Storage

Some products may be shipped at room temperature. This will not affect their stability or performance. Avoid repeated freeze/thaw cycles by aliquoting items into single-use fractions for storage at -20°C for up to 2 years. Keep all reagents on ice when not in storage.

Guarantee

This product is guaranteed for 12 months from date of receipt.

This product is for research use only and is not for use in diagnostic procedures.

Application Key

  • ChIP = Chromatin Immunoprecipitation;
  • ChIP = ChIP Sequencing;
  • CUT&RUN = Cleavage Under Targets and Release Using Nuclease;
  • CUT&Tag = Cleavage Under Targets and Tagmentation;
  • DB = Dot Blot;
  • ELISA = Enzyme-linked Immunosorbent Assay;
  • EMSA = Electrophoretic Mobility Shift Assay
  • FC = Flow Cytometry;
  • ICC = Immunocytochemistry;
  • IF = Immunofluorescence;
  • IHC = Immunohistochemistry;
  • IP = Immunoprecipitation;
  • MeDIP = Methyl-DNA Immunoprecipitation;
  • NEU = Neutralizing;
  • WB = Western Blotting