Description
General description
11kDa
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 H4 (Lys12) set includes the Acetyl-Histone H4 (Lys12) antibody, a negative control normal rabbit IgG, and qPCR primers which amplify a 213 bp region of human GAPDH coding region. The Acetyl-Histone H4 (Lys12) and negative controls are supplied in a scalable "per ChIP" reaction size and can be used to functionally validate the precipitation of Acetyl-Histone H4 (Lys12) associated chromatin.
Immunogen
Epitope: Unknown
Peptide corresponding to Histone H4 containing the sequence [GLG-AcK-GGA] on which Lys12 is acetylated.
Application
Chromatin Immunoprecipitation:
Sonicated chromatin prepared from HeLa cells (1 X 10E6 cell equivalents per IP) were subjected to chromatin immunoprecipitation using either 2 µL of Negative Control Supernatant, or 2 µL of Anti-Acetyl-Histone H4 (Lys12) and the Magna ChIP® A Kit (Cat. # 17-610).
Successful immunoprecipitation of Acetyl-Histone H4 (Lys12)-associated DNA fragments was verified by qPCR using ChIP Primers, human GAPDH Coding Region as a positive locus, and a gene desert region as a negative locus. (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.
Lysates from HeLa cells untreated or sodium butyrate treated (Lanes 1 and 2 respectively) were resolved probed with anti-acetyl-Histone H4 (Lys12) (1:1,000). Arrow indicates Acetyl-Histone H4 (Lys12).
Arrow indicates Acetyl-Histone H4 (Lys12) (~11 kDa) (Figure 3).
Research Category
Epigenetics & Nuclear Function
Research Sub Category
Histones
Chromatin Biology
This ChIPAb+ Acetyl-Histone H4 (Lys12) -ChIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.
Biochem/physiol Actions
This antibody detects histone H4 acetylated at Lys12.
Wide range of cross-reactivity expected based on sequence homology.
Packaging
25 assays per set. Recommended use: 2 μL of antibody per chromatin immunoprecipitation (dependent upon biological context).
Physical form
Anti-Acetyl-Histone H4 (Lys12) (rabbit monoclonal), . One vial containing 50 µL of rabbit monoclonal IgG cell culture supernatant in 0.1% sodium azide. Store at -20°C.
Negative Control Supernatant, . One vial containing 100 µL of cultured supernatant in 0.05% sodium azide. Store at -20°C.
ChIP Primers, human GAPDH coding region,. One vial containing 75 μL of 5 μM each primer specific for human GAPDH coding region.
Store at -20°C.
FOR: GGC TCC CAC CTT TCT CAT CC
REV: GGC CAT CCA CAG TCT TCT GG
Format: Purified
Protein A 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.
Analysis Note
Chromatin Immunoprecipitation:
Sonicated chromatin prepared from HeLa S3 cells (1 X 10E6 cell equivalents per IP) were subjected to chromatin immunoprecipitation using either 2 µL of Negative Control Supernatant, or 2 µL of Anti-Acetyl-Histone H4 (Lys12) and the Magna ChIP® A Kit (Cat. # 17-610).
Successful immunoprecipitation of Acetyl-Histone H4 (Lys12)-associated DNA fragments was verified by qPCR using ChIP Primers, human GAPDH 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 negative control supernatant and primers specific for human GAPDH coding region.
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.
