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Analysis Note

ControlIncludes normal rabbit IgG and primers specific for human cyclin B1 promoter region.

Application

This ChIPAb+ Acetyl-Histone H3 (Lys56) -ChIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.

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.

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.

Legal Information

MAGNA CHIP is a registered trademark of Merck KGaA, Darmstadt, Germany

UPSTATE is a registered trademark of Merck KGaA, Darmstadt, Germany

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'

Quality

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

Target description

~17 kDa observed