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Analysis Note

ControlIncludes negative control normal rabbit IgG and primers specific for human RPL10 promoter.

Application

Research Sub CategoryHistones

This ChIPAb+ Acetyl-Histone H4 (Lys16) -ChIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.

Research CategoryEpigenetics & Nuclear Function

Chromatin Immunoprecipitation:
Representative lot data.
Sonicated chromatin prepared from HeLa cells (1 X 10E6 cell equivalents per IP) were subjected to chromatin immunoprecipitation using either 2 µg of Normal rabbit IgG or 2 µL of Anti-acetyl-Histone H4 (Lys16) and the Magna ChIP™ A Kit (Cat. # 17-610).
Successful immunoprecipitation of acetyl-Histone H4 (Lys16) associated DNA fragments was verified by qPCR using ChIP Primers, RPL10 Promoter as a positive locus, and β-Globin primers as a negative locus. (Please see figures). 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.
Recombinant Histone H4 (Cat # 14-697) (lane1) and HeLa acid extract (lane 2) was resolved by electrophoresis, transferred to PVDF and probed with anti-Acetyl Histone H4 (Lys16) (0.1 µg/mL).
Proteins were visualized using a donkey anti-rabbit secondary antibody conjugated to HRP and a chemiluminescence detection system.
Arrow indicates Acetyl Histone H4 (Lys16) (~10 kDa) (Please see figures).

Multiplexing (Luminex™):
Representative lot data.
This antibody specifically recognizes histone H4 acetylated on Lys16 by Luminex assay (Please see figures).

Peptide Inhibition Assay:
Representative lot data.
This antibody peptide blocked on HeLa cell extracts (Please see figures).

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.

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 H4 (Lys16) set includes the Acetyl-Histone H4 (Lys16) antibody, a negative control normal rabbit IgG, and qPCR primers which amplify a 178 bp region of human RPL10 promoter. The Acetyl-Histone H4 (Lys16) 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 (Lys16)-associated chromatin.

Immunogen

KLH-conjugated Histone H4 corresponding to the N-terminus region at and around acetylated Lys16.

Epitope: Acetylated H4 Lys16

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: ~2 µL of antibody per chromatin immunoprecipitation (dependent upon biological context).

Physical form

Affinity purified

Anti-Acetyl-Histone H4 (Lys16) (rabbit polyclonal). One vial containing 50 µL of affinity purified rabbit polyclonal serum in buffer containing 0.1 M Tris-Glycine (pH 7.4) 150 mM NaCl with 0.05% sodium azide. Store at -20°C.

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, RPL10 promoter. One vial containing 75 µL of 5 µM of each primer specific for the promoter region of human RPL10. Store at -20°C.
FOR: ACC CGT CTT CGA CAG GAC T
REV: GGA ACG GAA GAC GAG AAC AG

Format: Purified

Quality

Chromatin Immunoprecipitation:
Sonicated chromatin prepared from HeLa cells (1 X 10E6 cell equivalents per IP) were subjected to chromatin immunoprecipitation using either 2 µg of Normal Rabbit IgG or 2 µL of Anti-acetyl-Histone H4 (Lys16) and the Magna ChIP^® A Kit (Cat. # 17-610).
Successful immunoprecipitation of acetyl-Histone H4 (Lys16) associated DNA fragments was verified by qPCR using ChIP Primers, RPL10 Promoter (Please see figures).
Please refer to the EZ-Magna ChIP™ A (Cat. # 17-408) or EZ-ChIP™ (Cat. # 17-371) protocol for experimental details.

Specificity

Broad species cross-reactivity is expected based on sequence homology.

This antibody detects histone H4 acetylated at Lys16.

Storage and Stability

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.

Target description

~10 kDa observed