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.
The HDAC1 antibody is made against a peptide corresponding to amino acids 467-482 of mouse HDAC1.
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).
Epigenetics & Nuclear Function
Research Sub Category
This ChIPAb+ HDAC1 -ChIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.
10 assays per kit, ~4μL per chromatin immunoprecipitation
Routinely evaluated by chromatin immunoprecipitation on U2OS cells and immunoblot on 3T3/A31 nuclear lysate.
Anti-HDAC1 (mouse monoclonal IgG1,supernatant). One vial containing 40 μL of culture supernatant with 0.05% sodium azide. Store at -20°C.
Negative ChIP Control Supernatant. One vial containing 40 uL of mouse IgG containing supernatant with 0.05% sodium azide. Store at -20°C.
ChIP Primers p21. One vial containing 75 μL of 5 μM of each primer specific for a region of the human p21 (WAF1/CIP1/CDKN1A) promoter. Store at -20°C.
P21 Forward: CCC ACA GCA GAG GAG AAA GAA
P21 Reverse: CTG GAA ATC TCT GCC CAG ACA
Stable for 1 year at -20°C from date of receipt
Included negative control antibody supernatant mouse IgG and control primers specific for human p21 (WAF1/CIP1/CDKN1A) promoter.
MAGNA CHIP is a registered trademark of Merck KGaA, Darmstadt, Germany
UPSTATE is a registered trademark of Merck KGaA, Darmstadt, Germany
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.