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Key Specifications Table
| Species Reactivity | Key Applications |
|---|---|
| H, B, M, R, Ht | WB, ChIP |
| Description | |
|---|---|
| Catalogue Number | 17-10237 |
| Trade Name |
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| Description | ChIPAb+™ HDAC2 - ChIP Validated Antibody and Primer Set |
| Overview | All ChIPAb+ antibodies are individually validated for chromatin precipitation, every lot, every time. Each ChIPAb+ antibody set includes control primers (tested every lot by qPCR) to biologically validate your IP results in a locus-specific context. The qPCR protocol and primer sequences are provided, allowing researchers to validate ChIP protocols when using our antibody in their chromatin context. Each set also includes a negative control antibody to ensure specificity of the ChIP reaction. The ChIPAb+ HDAC2 set includes the HDAC2 antibody, a Normal Mouse IgG, and control primers which amplify a 92 bp region of ChIP Primers, VWF promoter. The HDAC2 and negative controls are supplied in a scalable "per ChIP" reaction size and can be used to functionally validate the precipitation of HDAC2 -associated chromatin. |
| Alternate Names |
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| Background Information | Histone deacetylase 2 (HDAC2), or transcriptional regulator homolog RPD3 L1, is highly homologous to the yeast transcription factor RPD3 (reduced potassium dependency 3) gene. As in yeast, human HDA2 is likely to be involved in regulating chromatin structure during transcription. It has been implicated to associate with YY1, a mammalian zinc-finger transcription factor, which negatively regulates transcription by tethering RPD3 to DNA as a cofactor. This process is highly conserved from yeast to human. |
| Product Information | |
|---|---|
| Format | Purified |
| Control |
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| Presentation | Anti-HDAC2 (mouse monoclonal). One vial containing 50 µg of purified mouse monoclonal IgG in buffer containing 70% storage buffer (0.1M Tris-glycine, pH 7.4, 0.15 M NaCl, 0.05% sodium azide) and 30% glycerol. Store at -20° C. Concentration: 0.94 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, VWF promoter. One vial containing 75 μL of 5 μM of each primer specific for the promoter region of human von Willebrand factor. Store at -20°C. FOR: GCT GAG AGC ATG GCC TAG GGT GGT GGG CGG CAC REV: CCC CTG CAA ATG AGG GCT GCG GCT ATC TCC AAG |
| Quality Level | MQ100 |
| Applications | |
|---|---|
| Application | This ChIPAb+ HDAC2 -ChIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers. |
| Key Applications |
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| Application Notes | 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) was subjected to chromatin immunoprecipitation using using 2 µg of either Normal Mouse IgG or 2 µg of Anti-HDAC2 and the Magna ChIP™ A/G Kit (Cat. # 17-10085). Successful immunoprecipitation of HDAC2 associated DNA fragments was verified by qPCR using ChIP Primers, VWF promoter. Data is presented as percent input of each IP sample relative to input chromatin for each amplicon and ChIP sample as indicated. (Figure 2). 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 resolved by electrophoresis, transferred to nitrocellulose and probed with anti-HDAC2 (0.5 μg/mL). Proteins were visualized using a goat anti-mouse secondary antibody conjugated to HRP and a chemiluminescence detection system. Arrow indicates HDAC2 (~60 kDa). (Figure 3). Immunoprecipitation and HDAC Assay: Representative lot data. 3 μL of a previous lot was used to immunoprecipitate HDAC activity from HeLa nuclear extract, which was then measured using the HDAC Assay Kit (Fluorometric Detection) (Catalog # 17-356) (Figure 4). |
| Biological Information | |
|---|---|
| Immunogen | KLH-conjugated, synthetic peptide (CEKTDTKGTKSEQLSNP) corresponding to human HDAC2 at the C-terminus. |
| Epitope | C-terminus |
| Concentration | Please refer to the Certificate of Analysis for the lot-specific concentration. |
| Host | Mouse |
| Specificity | This antibody recognizes HDAC2 at the C-terminus. |
| Isotype | IgG1 |
| Species Reactivity |
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| Species Reactivity Note | Human, mouse, bovine and hamster. Predicted to cross-react with rat based on sequence homology. |
| Antibody Type | Monoclonal Antibody |
| Entrez Gene Number |
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| Entrez Gene Summary | This gene product belongs to the histone deacetylase family. Histone deacetylases act via the formation of large multiprotein complexes and are responsible for the deacetylation of lysine residues on the N-terminal region of the core histones (H2A, H2B, H3 and H4). This protein also forms transcriptional repressor complexes by associating with many different proteins, including YY1, a mammalian zinc-finger transcription factor. Thus it plays an important role in transcriptional regulation, cell cycle progression and developmental events. |
| Gene Symbol |
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| Purification Method | Protein G Purified |
| UniProt Number |
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| UniProt Summary | FUNCTION: SwissProt: Q92769 # Forms transcriptional repressor complexes by associating with MAD, SIN3, YY1 and N-COR. Interacts in the late S-phase of DNA-replication with DNMT1 in the other transcriptional repressor complex composed of DNMT1, DMAP1, PCNA, CAF1. SIZE: 488 amino acids; 55364 Da SUBUNIT: Interacts with the non-histone region of H2AFY (By similarity). Part of the core histone deacetylase (HDAC) complex composed of HDAC1, HDAC2, RBBP4 and RBBP7. The core complex associates with MTA2, MBD3, MTA1L1, CHD3 and CHD4 to form the nucleosome remodeling and histone deacetylation (NuRD) complex, or with SIN3, SAP18 and SAP30 to form the SIN3 HDAC complex. Component of a BHC histone deacetylase complex that contains HDAC1, HDAC2, HMG20B/BRAF35, AOF2/LSD1, RCOR1/CoREST and PHF21A/BHC80. The BHC complex may also contain ZMYM2, ZNF217, ZMYM3, GSE1 and GTF2I. Part of a complex containing the core histones H2A, H2B, H3 and H4, DEK and unphosphorylated DAXX. Part of a complex containing ATR and CHD4. Forms a heterologous complex at least with YY1. Interacts with ATR, DNMT1, MINT, HDAC7, HDAC10, HCFC1, NRIP1, MJD2A/JHDM3A, SAP30, SETDB1 and SUV39H1. Interacts with the non-histone region of H2AFY. Interacts with PELP1. Component of a mSin3A corepressor complex that contains SIN3A, SAP130, SUDS3/SAP45, ARID4B/SAP180, HDAC1 and HDAC2. Interacts with CBFA2T3. Interacts with SAP30L. SUBCELLULAR LOCATION: Nucleus.TISSUE SPECIFICITY: Widely expressed; lower levels in brain and lung. SIMILARITY: SwissProt: Q92769 ## Belongs to the histone deacetylase family. Type 1 subfamily. |
| Molecular Weight | ~60 kDa |
| Product Usage Statements | |
|---|---|
| Quality Assurance | Chromatin Immunoprecipitation: Representative lot data. Sonicated chromatin prepared from HEK293T cells (5 X 10E6 cell equivalents per IP) were subjected to chromatin immunoprecipitation using 2 µg of either Normal Mouse IgG, or 2 µg of Anti-HDAC2 and the Magna ChIP™ A/G Kit (Cat. # 17-10085). Successful immunoprecipitation of HDAC2 associated DNA fragments was verified by qPCR using ChIP Primers, VWF promoter (Figure 1). Please refer to the EZ-Magna ChIP™ A (Cat. # 17-408) or EZ-ChIP™ (Cat. # 17-371) protocol for experimental details |
| Usage Statement |
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| Storage and Shipping Information | |
|---|---|
| Storage Conditions | 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. |
| Packaging Information | |
|---|---|
| Material Size | 25 assays |
| Material Package | 25 assays per set. Recommended use: ~2 μg of antibody per chromatin immunoprecipitation (dependent upon biological context). |