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17-680

Sigma-Aldrich

ChIPAb+ Monomethyl-Histone H3 (Lys9) - ChIP Validated Antibody and Primer Set

clone CMA306, from mouse, purified by using protein G

Synonym(s):
H3K9me1, Histone H3 (mono methyl K9)
eCl@ss:
32160702

Quality Level

biological source

mouse

antibody form

purified immunoglobulin

clone

CMA306, monoclonal

purified by

using protein G

species reactivity

human, vertebrates

manufacturer/tradename

ChIPAb+
Upstate®

application(s)

ChIP: suitable (ChIP-seq)
immunoprecipitation (IP): suitable
western blot: suitable

isotype

IgG

NCBI accession no.

UniProt accession no.

shipped in

dry ice

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+ Monomethyl-Histone H3 (Lys9) set includes the Anti-monomethyl-Histone H3 (Lys9) antibody, a negative control antibody (purified Mouse IgG), and qPCR primers in the GAPDH coding region, amplifying a 213 base pair PCR product. The monomethyl-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 monomethyl-histone H3 (Lys9) associated chromatin.

Specificity

The immunogen sequence is identical in a wide range of animal and plant species.
Recognizes histone H3, Mr 17 kDa, monomethylated at Lys9.

Immunogen

Epitope: a.a. 1-18
The monomethyl-histone H3 (Lys9) purified antibody is made against a synthetic peptide (monomethylated at Lys9) corresponding to amino acids 1-18 of Histone H3.

Application

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-monomethyl-Histone H3 (Lys9) antibody and the Magna ChIP G (Cat. #17-611) Kit. Successful immunoprecipitation of monomethyl-histone H3 (Lys9) associated DNA fragments was verified by qPCR using GAPDH Coding region primers versus Control Primers directed against the GAPDH promoter region (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.

Western Blot Analysis:
Representative data of previous lot. HeLa acid extract (Lane 1) was resolved by electrophoresis, transferred to PVDF membrane and probed with Anti-monomethyl Histone H3 (Lys9) (0.5 μg/mL).
Proteins were visualized using a goat anti-mouse secondary antibody conjugated to HRP (Cat. #AP124P) and a chemiluminescence detection system (Please see figures).
This ChIPAb+ Validated Antibody & Primer Set conveniently includes the antibody, matched IgG negative control antibody & set control PCR primers that detect a known positive locus.
Research Category
Epigenetics & Nuclear Function
Research Sub Category
Chromatin Biology

Packaging

25 assays per kit, ~2μg per chromatin immunoprecipitation

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-monomethyl-Histone H3 (Lys9) antibody and the Magna ChIP G (Cat. #17-611) Kit. Successful immunoprecipitation of monomethyl-histone H3 (Lys9) associated DNA fragments was verified by qPCR using ChIP Primers GAPDH Coding region (Please see figures).
Please refer to the EZ-Magna G ChIP (Cat. #17-409) or EZ-ChIP (Cat. #17-371) protocol for experimental details.

Target description

Monomethyl-histone H3 at ~17 kDa

Physical form

Anti-monomethyl-Histone H3 (Lys9) (mouse monoclonal IgG2aқ, clone CMA306). One vial containing 50 μg of protein G purified antibody in 50 μL PBS containing 0.05% sodium azide. Store at -20°C.

Normal Mouse IgG. Two vials containing 25 μg purified Mouse IgG in 25 μL storage buffer containing 0.1% sodium azide. Store at -20°C.

ChIP Primers GAPDH Coding region. One vial containing 75 μL of 5 μM of each primer specific for a region of the human GAPDH coding region. Store at -20°C.
FOR: GGC TCC CAC CTT TCT CAT CC
REV: GGC CAT CCA CAG TCT TCT GG

Storage and Stability

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

Analysis Note

Control
Included negative control mouse IgG antibody and control primers specific for human GAPDH coding region.

Legal Information

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.

Certificate of Analysis

Certificate of Origin

Jane Ding et al.
Cell metabolism, 18(6), 896-907 (2013-12-10)
Increased activation of the serine-glycine biosynthetic pathway is an integral part of cancer metabolism that drives macromolecule synthesis needed for cell proliferation. Whether this pathway is under epigenetic control is unknown. Here we show that the histone H3 lysine 9...
Matthew J Harms et al.
Cell metabolism, 19(4), 593-604 (2014-04-08)
Prdm16 is a transcription factor that regulates the thermogenic gene program in brown and beige adipocytes. However, whether Prdm16 is required for the development or physiological function of brown adipose tissue (BAT) in vivo has been unclear. By analyzing mice...
Xian Li et al.
Frontiers in veterinary science, 7, 231-231 (2020-06-06)
Liver is the place where cholesterol is synthesized, transported, secreted, and transformed, thus liver takes an irreplaceable role in cholesterol homeostasis. Hepatic cholesterol metabolism differs between breeds, yet the molecular mechanism is unclear. In this study Large White (LW) and...
Liver-specific knockout of histone methyltransferase G9a impairs liver maturation and dysregulates inflammatory, cytoprotective, and drug-processing genes.
Lu, et al.
Xenobiotica, 49, 740-752 (2020)
Jingjing Meng et al.
Plant physiology, 177(2), 652-670 (2018-03-25)
DNA and histone methylation coregulate heterochromatin formation and gene silencing in animals and plants. To identify factors involved in maintaining gene silencing, we conducted a forward genetic screen for mutants that release the silenced transgene Pro35S::NEOMYCIN PHOSPHOTRANSFERASE II in the...

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