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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)

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About This Item

UNSPSC Code:
12352203
eCl@ss:
32160702
NACRES:
NA.52

biological source

mouse

Quality Level

antibody form

purified immunoglobulin

clone

CMA306, monoclonal

purified by

using protein G

species reactivity

vertebrates, human

manufacturer/tradename

ChIPAb+
Upstate®

technique(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

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

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).
Research Category
Epigenetics & Nuclear Function
Research Sub Category
Chromatin Biology
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.

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.

Storage Class Code

10 - Combustible liquids


Certificates of Analysis (COA)

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