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Key Documents

17-681

Sigma-Aldrich

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

clone CMA307, from mouse

Synonyme(s) :

H3K9me2, Histone H3 (di methyl K9)

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About This Item

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

Source biologique

mouse

Niveau de qualité

Clone

CMA307, monoclonal

Espèces réactives

human, vertebrates

Fabricant/nom de marque

ChIPAb+
Upstate®

Technique(s)

ChIP: suitable
western blot: suitable

Isotype

IgG

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+ 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.

Spécificité

Recognizes histone H3, Mr 17 kDa, dimethylated at lysine 9.
This peptide sequence is identical in a wide range of animal and plant species.

Immunogène

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.

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-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 Category
Epigenetics & Nuclear Function
Research Sub Category
Chromatin Biology
This ChIPAb+ Dimethyl-Histone H3 (Lys9) -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 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.

Description de la cible

Dimethyl-histone H3 at ~17 kDa

Forme physique

Anti-dimethyl-Histone H3 (Lys9) (mouse monoclonal IgG1қ, Clone CMA307). 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.

ChIP Primers B-Globin. One vial containing 75 μL of 5 μM of each primer specific for human β-globin.
Store at -20°C.
FOR: AGG ACA GGT ACG GCT GTC ATC
REV: TTT ATG CCC AGC CCT GGC TC
Format: Purified

Stockage et stabilité

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

Remarque sur l'analyse

Control
Included negative control mouse IgG antibody and control primers specific for human β-globin 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


Certificats d'analyse (COA)

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

Emanuela Sani et al.
Genome biology, 14(6), R59-R59 (2013-06-19)
In arid and semi-arid environments, drought and soil salinity usually occur at the beginning and end of a plant's life cycle, offering a natural opportunity for the priming of young plants to enhance stress tolerance in mature plants. Chromatin marks
Marcela A Bennesch et al.
Nucleic acids research, 44(18), 8655-8670 (2016-06-22)
The estrogen receptor α (ERα) is a transcription factor that can be directly activated by estrogen or indirectly by other signaling pathways. We previously reported that activation of the unliganded ERα by cAMP is mediated by phosphorylation of the transcriptional
Alba Fernández-Sánchez et al.
Epigenetics, 8(1), 66-78 (2012-12-14)
The human activating receptor NKG2D is mainly expressed by NK, NKT, γδ T and CD8(+) T cells and, under certain conditions, by CD4(+) T cells. This receptor recognizes a diverse family of ligands (MICA, MICB and ULBPs 1-6) leading to
Xin Lin et al.
Plant methods, 8(1), 48-48 (2012-12-12)
In this report we describe a chromatin immunoprecipitation (ChIP) protocol for two fully sequenced model diatom species Phaeodactylum tricornutum and Thalassiosira pseudonana. This protocol allows the extraction of satisfactory amounts of chromatin and gives reproducible results. We coupled the ChIP
Rui Wang et al.
Journal of the American Chemical Society, 135(3), 1048-1056 (2012-12-19)
Protein methyltransferases (PMTs) have emerged as important epigenetic regulators in myriad biological processes in both normal physiology and disease conditions. However, elucidating PMT-regulated epigenetic processes has been hampered by ambiguous knowledge about in vivo activities of individual PMTs particularly because

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