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Key Documents

17-678

Sigma-Aldrich

ChIPAb+ Trimethyl-Histone H3 (Lys4) - ChIP Validated Antibody and Primer Set

clone CMA304, from mouse, purified by using protein G

Synonyme(s) :

H3K4me3, Histone H3 (tri methyl K4)

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About This Item

Code UNSPSC :
12352203
eCl@ss :
32160702
Nomenclature NACRES :
NA.32

Source biologique

mouse

Niveau de qualité

Clone

CMA304, monoclonal

Produit purifié par

using protein G

Espèces réactives

human, vertebrates

Fabricant/nom de marque

ChIPAb+
Upstate®

Technique(s)

ChIP: suitable (ChIP-seq)
immunocytochemistry: suitable
western blot: suitable

Isotype

IgG1κ

Numéro d'accès NCBI

Numéro d'accès UniProt

Conditions d'expédition

dry ice

Description générale

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 H3 (Lys4) set includes the Anti-trimethyl-Histone H3 (Lys4) antibody, a negative control antibody (purified Mouse IgG), and qPCR primers which amplify a 166 bp region within the promoter of the human GAPDH gene. The trimethyl-Histone H3 (Lys4) 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 H3 (Lys4)-associated chromatin.
The previously assigned protein identifier Q66I33 has been merged into P84243. Full details can be found on the UniProt database.

Spécificité

Recognizes histone H3, Mr 17 kDa, trimethylated at Lys4.
The immunogen sequence is identical in a wide range of animal and plant species.

Immunogène

Epitope: a.a. 1-12
The trimethyl-histone H3 (Lys4) purified antibody is made against a synthetic peptide (trimethylated at Lys4) corresponding to amino acids 1-12 of human histone H3.

Application

Chromatin Immunoprecipitation:
Sonicated chromatin prepared from untreated or colcemid treated 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-trimethyl-Histone H3 (Lys4) antibody and the Magna ChIP G Kit (Cat. #17-611). Successful immunoprecipitation of trimethyl-Histone H3 (Lys4) associated DNA fragments was verified by qPCR using primers ampliflying a region of the human B-Globin promoter or using GAPDH promoter 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.

Western Blot Analysis:
Representative lot data.
HeLa acid extract were resolved by electrophoresis, transferred to PVDF membrane and probed with Anti-trimethyl Histone H3 (Lys4), clone CMA304 at 1 μg/ml (lane 1) or 0.5 μg/ml (lane 2). 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+ Trimethyl-Histone H3 (Lys4) -ChIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.

Conditionnement

25 assays per kit, ~2μg per chromatin immunoprecipitation

Qualité

Chromatin Immunoprecipitation:
Sonicated chromatin prepared from colcemid-reated HeLa cells (1 X 106 cell equivalents per IP) were subjected to chromatin immunoprecipitation using 2 µg of either normal mouse IgG or Anti-trimethyl-Histone H3 (Lys4) antibody and the Magna ChIP G Kit (Cat. #17-611). Successful immuno-precipitation of trimethyl-histone H3 (Lys4) associated DNA fragments was verified by qPCR using Control Primers (Please see figures).
Please refer to the EZ-Magna G ChIP (Cat. #17-409) or EZ-ChIP (Cat. #17-371) protocol for experimental details.

Description de la cible

Trimethyl histone H3 at ~17 kDa

Forme physique

Anti-trimethyl-Histone H3 (Lys4) (mouse monoclonal IgG1, Clone CMA304). One vial containing 50 μg of protein G purified antibody in 50 μL PBS containing 0.05% sodium. 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.

Control Primers. One vial containing 75 μL of 5 μM of each primer specific for the promoter region of GAPDH. Store at -20°C.
FOR: TAC TAG CGG TTT TAC GGG CG
REV: TCG AAC AGG AGG AGC AGA GAG CGA

Stockage et stabilité

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

Remarque sur l'analyse

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

Informations légales

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

Clause de non-responsabilité

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.

Code de la classe de stockage

10 - Combustible liquids


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Recherchez un Certificats d'analyse (COA) en saisissant le numéro de lot du produit. Les numéros de lot figurent sur l'étiquette du produit après les mots "Lot" ou "Batch".

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Consulter la Bibliothèque de documents

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Liss, A; Ooi, CH; Zjablovskaja, P; Benoukraf, T; Radomska, HS; Ju, C; Wu, M; Balastik et al.
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Jonathan C Irish et al.
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Judy A Brusslan et al.
Plant physiology, 168(4), 1246-1261 (2015-03-25)
The genome-wide abundance of two histone modifications, H3K4me3 and H3K9ac (both associated with actively expressed genes), was monitored in Arabidopsis (Arabidopsis thaliana) leaves at different time points during developmental senescence along with expression in the form of RNA sequencing data.
Judy A Brusslan et al.
PloS one, 7(3), e33151-e33151 (2012-03-20)
Leaf senescence is the orderly dismantling of older tissue that allows recycling of nutrients to developing portions of the plant and is accompanied by major changes in gene expression. Histone modifications correlate to levels of gene expression, and this study
Liang Yan et al.
Molecular pharmacology, 92(2), 113-123 (2017-05-27)
CYP3A4 is one of the major drug-metabolizing enzymes in human and is responsible for the metabolism of 60% of clinically used drugs. Many drugs are able to induce the expression of CYP3A4, which usually causes drug-drug interactions and adverse drug

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