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Merck

17-625

Sigma-Aldrich

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

from rabbit

동의어(들):

H3K9me3, Histone H3 (tri methyl K9), Histone H3K9me3

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크기 선택

제품정보 (DICE 배송 시 비용 별도)

UNSPSC 코드:
12352203
eCl@ss:
32160702
NACRES:
NA.41

주요 문서

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기술 서비스
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기술 서비스
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도움 문의

생물학적 소스

rabbit

Quality Level

100

클론

polyclonal

종 반응성

human, mouse, vertebrates

제조업체/상표

ChIPAb+
Upstate®

기술

ChIP: suitable
dot blot: suitable
western blot: suitable

NCBI 수납 번호

NP_003484

UniProt 수납 번호

Q16695

배송 상태

dry ice

유전자 정보

human ... H3F3B(3021)

관련 카테고리

일반 설명

17kDa
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 (Lys9) set includes the Anti-trimethyl-Histone H3 (Lys9) antibody, a negative control antibody (purified rabbit IgG), and qPCR primers which amplify a 117 bp region within the 3’ end of the human ZNF554 gene. The trimethyl-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 trimethyl-histone H3 (Lys9) associated chromatin.
The methylation of histones can occur on two different residues: arginine or lysine. Histone methylation can be associated with transcriptional activation or repression, depending on the methylated residue. Lysine 9 of histone H3 can be mono-, di- or trimethylated by different histone methyltransferases (HMTs) such as SuvH39H1 or G9a. This methylated lysine can be demethylated by histone demethylases as JMJD1A, LSD1 or JMJD2C. Methylation of this residue is mainly associated with transcriptional repression.

면역원

The trimethyl-histone H3 (Lys9) purified antibody is made against BSA-conjugated, synthetic peptide containing the sequence …AR[me3K]S… in which me3K corresponds to trimethyl lysine 9 of human Histone H3.

애플리케이션

Chromatin Immunoprecipitation:
Sonicated chromatin prepared from 2x106 HeLa cells were subjected to chromatin immunoprecipitation using 4 μg purified antibody or normal rabbit IgG and the Magna ChIP A kit (Cat. # 17-610).
Successful enrichment of trimethyl-Histone H3 (Lys9) associated DNA fragments was verified by qPCR using ChIP Primers GAPDH flanking the human GAPDH promoter and primers targeting the promoter of human MyoD.
Please refer to the EZ-Magna ChIP A (Cat. # 17-408) or EZ-ChIP (Cat. # 17-371) kit protocols for experimental details.

Western Blot Analysis:
Recombinant Histone H3 (Lane 1) and HeLa acid extract (Lane 2) were resolved by electrophoresis, transferred to nitrocellulose and probed with anti-trimethyl-Histone H3 (Lys9), (1 μg/mL). Proteins were visualized using a goat anti-rabbit secondary antibody conjugated to HRP and a chemiluminescence detection system (Please see figures).

Beadlyte Histone Peptide Specificity Assay:
0.5 μg/ml of purified anti-dimethyl-Histone H3 (Lys9) was incubated with a cocktail of microspheres conjugated to histone H3 peptides with the following modifications:
1. trimethyl-lysine 9
2. dimethyl-lysine 9
3. monomethyl-lysine 9
4. Unmodified H3
Unbound antibody was then removed by filtration. Peptide antibody complexes were incubated with a PE-conjugated anti-rabbit secondary antibody. Fluorescence was read on a Luminex 100 instrument. Median Fluorescence intensity (MFI) is plotted.
Trimethyl-Histone H3 (Lys9) ChIP validated antibody & primer set including the ChIP-grade antibody & the specific control PCR primers used for chromatin immunoprecipitation of H3K9Me3.

생화학적/생리학적 작용

Recognizes histone H3, Mr 17 kDa, trimethylated at lysine 9.
The immunogen sequence is identical in a wide range of animal and plant species, so broad cross-reactivity is expected.

물리적 형태

Anti-trimethyl-Histone H3 (Lys9) (rabbit polyclonal IgG). One vial containing 100 μg protein A purified IgG in 100 μL of 0.02 M Phosphate buffer, pH7.4, 0.25 M NaCl, 0.05% sodium azide with 30% glycerol. Store at -20°C.

Normal Rabbit IgG. One vial containing 125 μg of normal rabbit IgG in 125 μL storage buffer containing 0.1% sodium azide. Store at -20°C.

ChIP Primers, ZNF554. One vial containing 75 μL of 5 μM of each primer specific for ZNF554. Store at -20°C.
FOR: CGG GGA AAA GCC CTA TAA AT
REV: TCC ACA TTC ACT GCA TTC GT
Format: Purified

분석 메모

Chromatin Immunoprecipitation:
Sonicated Chromatin prepared from 3x106 NIH3T3 L1 cells were subjected to chromatin immunoprecipitation using 4 µg of either normal rabbit IgG or Anti-trimethyl-Histone H3 (Lys9) antibody and the Magna ChIP A kit (Cat. #17-610). Successful enrichment of trimethyl-histone H3 (Lys9)-associated DNA fragments was verified by qPCR using ChIP Primers ZNF554 (Please see figures).
Please refer to the EZ-Magna A ChIP (Cat. #17-408) or EZ-ChIP (Cat. #17-371) protocol for experimental details.
Control
Included negative control rabbit IgG antibody and control primers specific for ZNF554.

기타 정보

Anti-trimethyl Histone H3 (Lys9) polyclonal, 1 vial

ZNF554 primer set, 1 vial

법적 정보

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

Storage Class Code

10 - Combustible liquids

시험 성적서(COA)

제품의 로트/배치 번호를 입력하여 시험 성적서(COA)을 검색하십시오. 로트 및 배치 번호는 제품 라벨에 있는 ‘로트’ 또는 ‘배치’라는 용어 뒤에서 찾을 수 있습니다.

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Large intronic expansions of the triplet-repeat sequence (GAA.TTC) cause transcriptional repression of the Frataxin gene (FXN) leading to Friedreich's ataxia (FRDA). We previously found that GAA-triplet expansions stimulate heterochromatinization in vivo in transgenic mice. We report here using chromosome conformation
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Advance your Epigenetics Research

Epigenetics describes heritable changes in gene expression caused by non-genetic mechanisms. Epigenetic regulation allows a cell to vary its response based on its biological and environmental contexts. Epigenetic changes can effect transcriptional and post-transcriptional regulation via mechanisms such as histone modification, chromatin and nucleosome remodeling, DNA methylation, and small and non-coding RNA-mediated regulation. These mechanisms, in cooperation with transcription factors and other nucleic acid-binding proteins, regulate gene expression. Epigenetic mechanisms of gene regulation impacts diverse areas of research—from agriculture to human health. Common epigenetic assays such as chromatin immunoprecipitation (ChIP) and RNA immunoprecipitation (RIP) rely on high quality antibodies that recognize specific epigenetic modifications for accurate results. EMD Millipore offers over 100 ChIPAb+™ and RIPAb+™ validated antibody kits that are quality tested on ChIP/RIP assays and are conveniently provided with control qPCR primers and negative control antibodies to ensure first time ChIP/RIP success.

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