ChIPAb + HDAC1

Stock Code: 3586445
Manufacturer Part No: 17-608
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Analysis Note


ControlIncluded negative control antibody supernatant mouse IgG and control primers specific for human p21 (WAF1/CIP1/CDKN1A) promoter.


Application


This ChIPAb+ HDAC1 -ChIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.


Chromatin Immunoprecipitation Analysis:
Sonicated chromatin prepared from untreated or actinomycin D treated (30 ng/mL, 6 hrs.) U2OS cells (3 X 106 cell equivalents per IP) was subjected to chromatin immunoprecipitation using 4 µL of Mouse Anti-HDAC1 and the Magna ChIP® G (Cat. # 17-611) Kit. Immunoprecipitation of HDAC1 associated DNA fragments was verified by qPCR using ChIP Primers p21 flanking the human p21 promoter that contains an Sp1 binding site (Please see figures).
Fold Decrease is a ratio of normalized mean IP quantities extracted from standard curves derived from inputs of each chromatin sample. As previously published, HDAC1 association with this region of the p21 promoter decreases upon actinomycin D treatment.

Western Blot Analysis:
3T3/A31 cell lysate was resolved by electrophoresis, transferred to nitrocellulose and probed with anti-HDAC1 (1:2500). Proteins were visualized using a goat anti-mouse secondary antibody conjugated to HRP and a chemiluminescence detection system (Please see figures).


Research CategoryEpigenetics & Nuclear Function


Research Sub CategoryChromatin Biology


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+ HDAC1 set includes the HDAC1 antibody, the negative control antibody supernatant (mouse IgG), and qPCR primers flanking an Sp1 binding site in human p21 (WAF1/CIP1/CDKN1A) promoter, amplifying a 105 base pair PCR product. The HDAC1 and negative control antibodies are supplied in a scalable "per ChIP" reaction size and can be used to functionally validate the precipitation of HDAC1 associated chromatin.


Immunogen


The HDAC1 antibody is made against a peptide corresponding to amino acids 467-482 of mouse HDAC1.


Legal Information


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


UPSTATE is a registered trademark of Merck KGaA, Darmstadt, Germany


Packaging


10 assays per kit, ~4µL per chromatin immunoprecipitation


Physical form


Anti-HDAC1 (mouse monoclonal IgG1,supernatant)

Quality Level100
ManufacturerSIGMA-ALDRICH

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