dry ice
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 polyclonal set includes the HDAC1 antibody, a Normal Rabbit IgG, and control primers which amplify a 88 bp region of ChIP Primers p21. The HDAC1 and negative controls are supplied in a scalable "per ChIP" reaction size and can be used to functionally validate the precipitation of HDAC1-associated chromatin.
Immunogen
Synthetic peptide corresponding to amino acids 467-482 of human HDAC1.
Application
Research Sub Category
Histones
Research Category
Epigenetics & Nuclear Function
This ChIPAb+ HDAC1 -ChIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.
Chromatin Immunoprecipitation:
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 Rabbit IgG, or 2 µg of Anti-HDAC1 and the Magna ChIP® A Kit (Cat. # 17-610). Successful immunoprecipitation of HDAC1 associated DNA fragments was verified by qPCR using ChIP Primers, p21 . (Figure 2). Data is presented as percent input of each IP sample relative to input chromatin for each amplicon and ChIP sample as indicated.
Please refer to the EZ-Magna ChIP A (Cat. # 17-408) or EZ-ChIP (Cat. # 17-371) protocol for experimental details.
Western Blot Analysis:
0.5 µg/mL from a representative lot detected HDAC1 in HeLa cells. (Figure 3).
Immunofluorescence Analysis:
2 µg/mL from a representative lot detected HDAC1 in HeLa cells.
Packaging
25 assays per set. Recommended use: ~2 μg of antibody per chromatin immunoprecipitation (dependent upon biological context).
Quality
Chromatin Immunoprecipitation:
Sonicated chromatin prepared from U2OS cells (3 X 10E6 cell equivalents per IP) were subjected to chromatin immunoprecipitation using 2 µg of either Normal Rabbit IgG, or 2 µg of Anti-HDAC1 and the Magna ChIP® A Kit (Cat. # 17-610). Successful immunoprecipitation of HDAC1 associated DNA fragments was verified by qPCR using ChIP Primers, p21 flanking an Sp1 binding site in the human p21 promoter (Figure 1).
Please refer to the EZ-Magna ChIP A (Cat. # 17-408) or EZ-ChIP (Cat. # 17-371) protocol for experimental details.
