추천 제품
생물학적 소스
rabbit
Quality Level
클론
polyclonal
종 반응성
human, mouse, vertebrates
제조업체/상표
ChIPAb+
Upstate®
기술
ChIP: suitable
dot blot: suitable
western blot: suitable
NCBI 수납 번호
UniProt 수납 번호
배송 상태
dry ice
유전자 정보
human ... H3F3B(3021)
관련 카테고리
일반 설명
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 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.
특이성
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.
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.
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.
성분
Anti-trimethyl Histone H3 (Lys9) polyclonal, 1 vial
ZNF554 primer set, 1 vial
ZNF554 primer set, 1 vial
품질
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.
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.
표적 설명
17kDa
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
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
Control
Included negative control rabbit IgG antibody and control primers specific for ZNF554.
Included negative control rabbit IgG antibody and control primers specific for ZNF554.
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.
Dicer independent small RNAs associate with telomeric heterochromatin.
Cao, F; Li, X; Hiew, S; Brady, H; Liu, Y; Dou, Y
RNA null
Polycomb repressive complex 2 regulates normal hematopoietic stem cell function in a developmental-stage-specific manner.
Xie, H; Xu, J; Hsu, JH; Nguyen, M; Fujiwara, Y; Peng, C; Orkin, SH
Cell Stem Cell null
Alba Millanes-Romero et al.
Molecular cell, 52(5), 746-757 (2013-11-19)
Although heterochromatin is enriched with repressive traits, it is also actively transcribed, giving rise to large amounts of noncoding RNAs. Although these RNAs are responsible for the formation and maintenance of heterochromatin, little is known about how their transcription is
The many faces of histone lysine methylation.
Lachner, Monika and Jenuwein, Thomas
Current Opinion in Cell Biology, 14, 286-298 (2002)
Chelsea Herdman et al.
PLoS genetics, 13(7), e1006899-e1006899 (2017-07-18)
Transcription of the several hundred of mouse and human Ribosomal RNA (rRNA) genes accounts for the majority of RNA synthesis in the cell nucleus and is the determinant of cytoplasmic ribosome abundance, a key factor in regulating gene expression. The
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