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+ Acetyl-Histone H3 (Lys56) set includes the Acetyl-Histone H3 (Lys56) antibody, a Normal Rabbit IgG, and control primers which amplify a 152 bp region of the human cyclin B1 promoter. The Acetyl-Histone H3 (Lys56) and negative controls are supplied in a scalable "per ChIP" reaction size and can be used to functionally validate the precipitation of Acetyl-Histone H3 (Lys56) associated chromatin.
~17 kDa observed
Application
Chromatin Immunoprecipitation:
Sonicated chromatin prepared from HeLa cells (1x104, 1x105 and 1x106 cell equivalents per IP) were subjected to chromatin immunoprecipitation using 1 µL of either Normal Rabbit IgG (Part No. CS213233), or 1 µL Anti-Acetyl-Histone H3 (Lys56) (Part No.CS207337) and the Magna ChIP HiSens Kit (Cat. # 17-10460). Successful immunoprecipitation of acetyl Histone H3 (Lys56) associated DNA fragments was verified by qPCR using ChIP Primers, human cyclin B1 promoter (Part No. CS207338).
Please refer to the MagnaChIP HiSens (Cat. # 17-10460) or EZ-MagnaChIP HiSens (Cat. # 17-10461) protocol for experimental details.
Dot Blot (Specificity) Analysis: A representative lot detected acetyl-Histone H3 (Lys56) in an Absurance Histone H3 Antibody Specificity Array (Cat. No. 16-667).
Western Blotting Analysis:
Evaluated by Western Blotting in HeLa acid extract, sodium butyrate treated HeLa acid extract, colcemid treated HeLa acid extract, recombinant Histone H3, recombinant Histone H4, and recombinant Histone H2A.
A 1:400 dilution of this antibody detected acetyl Histone H3 (Lys56) in sodium butyrate treated HeLa acid extract.
This ChIPAb+ Acetyl-Histone H3 (Lys56) -ChIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.
Packaging
25 assays per set. Recommended use: 1 μL of antibody per chromatin immunoprecipitation (dependent upon biological context).
Physical form
Anti-acetyl-Histone H3 (Lys56) (Rabbit Polyclonal). One vial containing 25 µL purified rabbit polyclonal in buffer containing 0.1 M Tris-Glycine (pH7.4),150mM NaCl, and 0.05% sodium azide, before the addition of 30% glycerol. Store at -20°C.
Normal Rabbit IgG. One vial containing 25 µL of normal rabbit IgG in buffer containing 0.01M PBS, 0.1% sodium azide, pH7.1, before the addition of 30% glycerol. Store at -20°C.
ChIP Primers, human cyclin B1 promoter. One vial containing 75 μL of 5 μM of each primer specific for human cyclin B1 promoter (chr5: 68462295-68462447, hg19 build). Store at -20°C.
FOR: 5’ CCT TGG GAA ACT GGA CAA TC 3’
REV: 5’ TTT GCC AGG GTC ACA CAT TA 3’
Analysis Note
Chromatin Immunoprecipitation:
Sonicated chromatin prepared from HeLa cells (1x105 cell equivalents per IP) were subjected to chromatin immunoprecipitation using 1 µL of either Normal Rabbit IgG (Part No. CS213233), or 1 µL Anti-Acetyl-Histone H3 (Lys56) (Part No.CS207337) and the Magna ChIP® HiSens Kit (Cat. # 17-10460). Successful immunoprecipitation of acetyl Histone H3 (Lys56) associated DNA fragments was verified by qPCR using ChIP Primers, human cyclin B1 promoter (Part No. CS207338).
Please refer to the MagnaChIP HiSens (Cat. # 17-10460) or EZ-MagnaChIP HiSens (Cat. # 17-10461) protocol for experimental details
Control
Includes normal rabbit IgG and primers specific for human cyclin B1 promoter region.
Legal Information
MAGNA CHIP is a registered trademark of Merck KGaA, Darmstadt, Germany
UPSTATE is a registered trademark of Merck KGaA, Darmstadt, Germany
