Description
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.
~62 kDa observed
Immunogen
Synthetic peptide corresponding to amino acids 467-482 of human HDAC1.
Application
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 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:
Representative lot data.
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.
Research Category
Epigenetics & Nuclear Function
Research Sub Category
Histones
This ChIPAb+ HDAC1 -ChIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.
Packaging
25 assays per set. Recommended use: ~2 μg of antibody per chromatin immunoprecipitation (dependent upon biological context).
Physical form
Anti-HDAC1 (rabbit polyclonal). One vial containing 50 µg of Anti-HDAC1 IgG in PBS with 1 mg/mL BSA and ≤0.1% sodium azide before the addition of glycerol to 30%. Store at -20°C.
Concentration: 0.7 mg/mL
Normal Rabbit IgG. One vial containing 125 µg of rabbit IgG in 125 µL of storage buffer containing 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.
FOR: CCC ACA GCA GAG GAG AAA GAA
REV: CTG GAA ATC TCT GCC CAG ACA
Format: Purified
Preparation Note
Stable for 1 year at -20°C from date of receipt. Handling Recommendations: Upon first thaw, and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance.
Note: Variability in freezer temperatures below -20°C may cause glycerol containing solutions to become frozen during storage.
Analysis Note
Chromatin Immunoprecipitation:
Representative lot data.
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.
Control
Includes normal rabbit IgG and primers specific for human p21.
Other Notes
Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.
Legal Information
MAGNA CHIP is a registered trademark of Merck KGaA, Darmstadt, Germany
UPSTATE is a registered trademark of Merck KGaA, Darmstadt, Germany
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.
