Not available outside of the UK & Ireland.

Analysis Note

ControlIncluded negative control mouse IgG antibody and control primers specific for human β-globin promoter.

Application

Research CategoryEpigenetics & Nuclear Function

Chromatin Immunoprecipitation:
Sonicated chromatin prepared from HeLa cells (1 X 10⁶ cell equivalents per IP) was subjected to chromatin immunoprecipitation using 2 µg of either a normal mouse IgG or Anti-dimethyl-Histone H3 (Lys9) antibody and the Magna ChIP G (Cat. #17-611) Kit. Successful immunoprecipitation of dimethyl-histone H3 (Lys9) associated DNA fragments was verified by qPCR using β-globin ChIP Primers versus Control Primers (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 G ChIP™ (Cat. #17-408) or EZ-ChIP™ (Cat. #17-371) protocol for experimental details.

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 Anti-dimethyl H3 (Lys9) at 2.0ug/mL (1:500) dilution. Proteins were visualized using a Donkey anti-mouse IgG conjugated to HRP and a chemiluminescence detection system.

Western Blot Analysis:
Recombinant Histone H3 (Lane 1) and HeLa acid extract (Lane 2) were resolved by electrophoresis, transferred to PVDF membrane and probed with Anti-dimethyl Histone H3 (Lys9), clone CMA307 (0.1 µg/ml). Proteins were visualized using a goat anti-mouse secondary antibody conjugated to HRP and a chemiluminescence detection system.

Multiplexing:
This antibody specifically recognizes histone H3 dimethylated on Lys9 by Luminex™ assay.

ELISA: This antibody has been shown by an outside laboratory to be suitable for ELISA.

Immunocytochemistry: This antibody has been shown by an outside laboratory to be suitable for immunocytochemistry.

Immunoprecipitation: This antibody has been shown by an outside laboratory to be suitable for immunoprecipitation.

Research Sub CategoryChromatin Biology

This ChIPAb+ Dimethyl-Histone H3 (Lys9) -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+ Dimethyl-Histone H3 (Lys9) set includes the Anti-dimethyl-Histone H3 (Lys9) antibody, a negative control antibody (purified Mouse IgG), and qPCR primers which amplify a 110 bp region within the promoter of the human β-globin gene. The dimethyl-histone H3 (Lys9) and negative control antibodies are supplied in a scalable "per ChIP" reaction size and can be used to functionally validate the precipitation of dimethyl-histone H3 (Lys9) associated chromatin.

Immunogen

Epitope: a.a. 1-18

The dimethyl-histone H3 (Lys9) purified antibody is made against a synthetic peptide (dimethylated at Lys9) corresponding to amino acids 1-18 of histone H3.

Legal Information

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

Packaging

25 assays per kit, ~2µg per chromatin immunoprecipitation

Physical form

Format: Purified

Quality

Chromatin Immunoprecipitation:
Sonicated chromatin prepared from HeLa cells (1 X 10⁶ cell equivalents per IP) were subjected to chromatin immunoprecipitation using 2 µg of either a normal mouse IgG or Anti-dimethyl-Histone H3 (Lys9) antibody and the Magna ChIP G (Cat. #17-611) Kit. Successful immunoprecipitation of dimethyl-histone H3 (Lys9) associated DNA fragments was verified by qPCR using ChIP Primers B-Globin (Please see figures).
Please refer to the EZ-Magna G ChIP™ (Cat. #17-409) or EZ-ChIP™ (Cat. #17-371) protocol for experimental details.

Specificity

Recognizes histone H3, Mr 17 kDa, dimethylated at lysine 9.

This peptide sequence is identical in a wide range of animal and plant species.

Storage and Stability

Stable for 1 year at -20°C from date of receipt.
Aliquot upon initial thaw, avoid freeze thaw cycles.

Target description

Dimethyl-histone H3 at ~17 kDa