ChIPAb+ HDAC1 - ChIP Validated Antibody and Primer Set
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REFERENCES
cAMP-responsive element modulator (CREM)α protein induces interleukin 17A expression and mediates epigenetic alterations at the interleukin-17A gene locus in patients with systemic lupus erythematosus.
Rauen, Thomas, et al. (2011) J. Biol. Chem., 286: 43437-46 (2011)
cAMP-responsive element modulator (CREM)α protein signaling mediates epigenetic remodeling of the human interleukin-2 gene: implications in systemic lupus erythematosus.
Hedrich, Christian M, et al. (2011) J. Biol. Chem., 286: 43429-36 (2011)
Species Reactivity Key Applications Host Format Antibody Type
H, M WB, ChIP Mouse Culture Supernatant Monoclonal Antibody
Description:
ChIPAb+ HDAC1 - ChIP Validated Antibody and Primer Set
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Trade Name:
Upstate (Millipore)
Product 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+ 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.
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Specificity:
HDAC1
Molecular Weight:
60 kDa
Immunogen:
The HDAC1 antibody is made against a peptide corresponding to amino acids 467-482 of mouse HDAC1.
Isotype:
IgG1
Species Reactivity:
Human
Mouse
Application Notes:
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).
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Control:
Included negative control antibody supernatant mouse IgG and control primers specific for human p21 (WAF1/CIP1/CDKN1A) promoter.
Quality Assurance:
Routinely evaluated by chromatin immunoprecipitation on U2OS cells and immunoblot on 3T3/A31 nuclear lysate.
Presentation:
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 super |