ChIPAb+ Trimethyl-Histone H4 (Lys20)

Stock Code: 3586467
Manufacturer Part No: 17-671

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Descriptions

Analysis Note

ControlIncludes negative control rabbit antibody and control primers specific for human β-globin promoter.

Application

Research CategoryEpigenetics & Nuclear Function

Chromatin Immunoprecipitation:
Sonicated chromatin prepared from HeLa cells (2 X 10⁶ cell equivalents per IP) was subjected to chromatin immunoprecipitation using 10 µL of either a negative control supernatant or Anti-Trimethyl-Histone H4 (Lys20) antibody and the Magna ChIP A Kit (Part No. 17-610). Successful immunoprecipitation of trimethylhistone H4 (Lys20) associated DNA fragments was verified by qPCR using β-globin ChIP Primers versus Control Primers corresponding to the GAPDH promoter (Please see figures). Data is presented as percent input of each IP sample relative to input chromatin, with immunoprecipitated DNA from negative control supernatant shown as (-) and trimethyl histone H4 (Lys20) shown as (+).
Please refer to the EZ-Magna ChIP™ A (Cat. # 17-408) or EZ-ChIP™ (Cat. # 17-371) protocol for experimental details.

Western Blot Analysis:
Acid extracts from HeLa cells untreated or pre-absorbed with 1 µM of trimethylated (Lys20) Histone H4 peptide (Lanes 1 and 2 respectively) were probed with anti-trimethyl-Histone H4 (Lys20) (1:1,000) (Please see figures).

Dot Blot Analysis: Absurance Histone H3 Antibody Specificity Array (Cat. No. 16-667) and Absurance Histone H2A, H2B, H4 Antibody Specificity Array (Cat. No. 16-665), which contain histone peptides with various modifications were probed with Cat. No 04-079, Anti-trimethyl-Histone H4 (Lys20) ; dilution 1:500. Proteins were visualized using a Donkey anti-rabbit IgG conjugated to HRP and a chemiluminescence detection system.

Research Sub CategoryChromatin BiologyHistones

This ChIPAb+ Trimethyl-Histone H4 (Lys20) -ChIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.

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+ Trimethyl-Histone H4 (Lys20) set includes the Anti-trimethyl-Histone H4 (Lys20) antibody, a negative control supernatant, and qPCR primers which amplify a 110 bp region within the promoter of the humanβ-globin gene. The trimethyl histone H4 (Lys20) and negative control antibodies are supplied in a scalable "per ChIP" reaction size and can be used to functionally validate the precipitation of trimethyl-histone H4 (Lys20) associated chromatin.