ChIPAb+ HDAC2; clone 3F3

Stock Code: 3586365
Manufacturer Part No: 17-10237
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Analysis Note


ControlIncludes normal mouse IgG and primers specific for human von Willebrand factor.


Application


Research Sub CategoryHistones


This ChIPAb+ HDAC2 -ChIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.


Research CategoryEpigenetics & Nuclear Function


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).



Disclaimer


Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.


General description


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.


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

Quality Level100
ManufacturerSIGMA-ALDRICH

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