一般說明
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+ Histone H2B set includes the Histone H2B antibody, a negative control rabbit IgG, and qPCR primers which amplify a 110 bp region of human β-globin promoter. The Histone H2B and negative controls are supplied in a scalable "per ChIP" reaction size and can be used to functionally validate the precipitation of Histone H2B-associated chromatin.
The ChIPAb+ Histone H2B set includes the Histone H2B antibody, a negative control rabbit IgG, and qPCR primers which amplify a 110 bp region of human β-globin promoter. The Histone H2B and negative controls are supplied in a scalable "per ChIP" reaction size and can be used to functionally validate the precipitation of Histone H2B-associated chromatin.
Histone H2B is one of the 5 main histone proteins involved in the structure of chromatin in eukaryotic cells. Featuring a main globular domain and a long N terminal tail H2A is involved with the structure of the nucleosomes of the ′beads on a string′ structure.
特異性
Broad species cross-reactivity expected based on sequence homology.
Others not tested.
Recognizes and is specific for Histone H2B, MW ~15 kDa. An additional band at ~23 kDa is detected in some samples, which likely corresponds to ubiquityl-Histone H2B.
免疫原
KLH-conjugated, synthetic peptide corresponding to amino acids 118-126 (CG-AVTKYTSSK) of human Histone H2B, with an N-terminal CG added for conjugation purposes.
應用
Chromatin Immunoprecipitation:
Sonicated chromatin prepared from HeLa cells (1 X 106 cell equivalents per IP) were subjected to chromatin immunoprecipitation using either 1 µL of Normal Rabbit IgG or 1 µL of Anti-Histone H2B and the Magna ChIP A Kit (Cat. # 17-610).
Successful immunoprecipitation of Histone H2B associated DNA fragments was verified by qPCR using ChIP Primers, human β-globin for a positive, and human GAPDH primers as a negative assay (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 ChIP A (Cat. # 17-408) or EZ-ChIP (Cat. # 17-371) protocol for experimental details.
Western Blot Analysis:
Representative lot data.
Recombinant Xenopus Histone H2B, chicken core histones and acid extract from either sodium butyrate or colcemid treated HeLa cells were resolved by electrophoresis, transferred to nitrocellulose and probed with anti-Histone H2B (1:5,000 dilution).
Proteins were visualized using a goat anti-rabbit secondary antibody conjugated to HRP and a chemiluminescence detection system (Please see figures).
Sonicated chromatin prepared from HeLa cells (1 X 106 cell equivalents per IP) were subjected to chromatin immunoprecipitation using either 1 µL of Normal Rabbit IgG or 1 µL of Anti-Histone H2B and the Magna ChIP A Kit (Cat. # 17-610).
Successful immunoprecipitation of Histone H2B associated DNA fragments was verified by qPCR using ChIP Primers, human β-globin for a positive, and human GAPDH primers as a negative assay (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 ChIP A (Cat. # 17-408) or EZ-ChIP (Cat. # 17-371) protocol for experimental details.
Western Blot Analysis:
Representative lot data.
Recombinant Xenopus Histone H2B, chicken core histones and acid extract from either sodium butyrate or colcemid treated HeLa cells were resolved by electrophoresis, transferred to nitrocellulose and probed with anti-Histone H2B (1:5,000 dilution).
Proteins were visualized using a goat anti-rabbit secondary antibody conjugated to HRP and a chemiluminescence detection system (Please see figures).
Research Category
Epigenetics & Nuclear Function
Epigenetics & Nuclear Function
Research Sub Category
Histones
Histones
The ChIPAb+ Histone H2B set includes the Histone H2B antibody, a negative control rabbit IgG & qPCR primers which amplify a 110 bp region of human β-globin promoter.
包裝
25 assays per set. Recommended use: ~1 μL of antibody per chromatin immunoprecipitation (dependent upon biological context).
品質
Chromatin Immunoprecipitation:
Sonicated chromatin prepared from HeLa cells (1 X 106 cell equivalents per IP) were subjected to chromatin immunoprecipitation using either 1 µL of Normal Rabbit IgG or 1 µL of Anti-Histone H2B and the Magna ChIP® A Kit (Cat. # 17-610).
Successful immunoprecipitation of Histone H2B associated DNA fragments was verified by qPCR using ChIP Primers, human β-globin (Please see figures).
Please refer to the EZ-Magna ChIP A (Cat. # 17-408) or EZ-ChIP (Cat. # 17-371) protocol for experimental details.
Sonicated chromatin prepared from HeLa cells (1 X 106 cell equivalents per IP) were subjected to chromatin immunoprecipitation using either 1 µL of Normal Rabbit IgG or 1 µL of Anti-Histone H2B and the Magna ChIP® A Kit (Cat. # 17-610).
Successful immunoprecipitation of Histone H2B associated DNA fragments was verified by qPCR using ChIP Primers, human β-globin (Please see figures).
Please refer to the EZ-Magna ChIP A (Cat. # 17-408) or EZ-ChIP (Cat. # 17-371) protocol for experimental details.
標靶描述
~15 kDa
外觀
Protein A purified
Anti-Histone H2B (rabbit polyclonal). One vial containing 25 µL of protein A purified rabbit polyclonal in buffer containing 0.02 M phosphate buffer, pH 7.6, 0.25 M NaCl, and 0.1% sodium azide, and 30% glycerol. Store at -20°C.
Normal Rabbit IgG. One vial containing 125 μg of purified rabbit IgG in 125 μL storage buffer. Store at -20°C.
ChIP Primers, β-globin. One vial containing 75 μL of 5 μM of each primer specific for the human β-globin promoter. Store at -20°C.
FOR: AGG ACA GGT ACG GCT GTC ATC
REV: TTT ATG CCC AGC CCT GGC TC
Normal Rabbit IgG. One vial containing 125 μg of purified rabbit IgG in 125 μL storage buffer. Store at -20°C.
ChIP Primers, β-globin. One vial containing 75 μL of 5 μM of each primer specific for the human β-globin promoter. Store at -20°C.
FOR: AGG ACA GGT ACG GCT GTC ATC
REV: TTT ATG CCC AGC CCT GGC TC
Format: Purified
儲存和穩定性
Stable for 1 year at -20°C from date of receipt. Handling Recommendations: Upon first thaw, and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance. Note: Variabillity in freezer temperatures below -20°C may cause glycerol containing solutions to become frozen during storage.
分析報告
Control
Includes negative control normal rabbit IgG antibody and primers specific for human β-globin.
Includes negative control normal rabbit IgG antibody and primers specific for human β-globin.
法律資訊
MAGNA CHIP is a registered trademark of Merck KGaA, Darmstadt, Germany
UPSTATE is a registered trademark of Merck KGaA, Darmstadt, Germany
免責聲明
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.
儲存類別代碼
10 - Combustible liquids
Jiwon Hwang et al.
mBio, 2(2), e00023-e00011 (2011-03-31)
Elongins B and C are members of complexes that increase the efficiency of transcriptional elongation by RNA polymerase II (RNAPII) and enhance the monoubiquitination of histone H2B, an epigenetic mark of actively transcribed genes. Here we show that, in addition
Shaun D McCullough et al.
Proceedings of the National Academy of Sciences of the United States of America, 109(52), 21319-21324 (2012-12-14)
Spinocerebellar ataxia type 7 (SCA7) is an autosomal-dominant neurodegenerative disorder that results from polyglutamine expansion of the ataxin-7 (ATXN7) protein. Remarkably, although mutant ATXN7 is expressed throughout the body, pathology is restricted primarily to the cerebellum and retina. One major
Xianjiang Lan et al.
Molecular and cellular biology, 35(10), 1777-1787 (2015-03-11)
Spinocerebellar ataxia type 7 (SCA7) is a debilitating neurodegenerative disease caused by expansion of a polyglutamine [poly(Q)] tract in ATXN7, a subunit of the deubiquitinase (DUB) module (DUBm) in the SAGA complex. The effects of ATXN7-poly(Q) on DUB activity are
Guang Yang et al.
Nucleic acids research, 48(6), 3001-3013 (2020-01-23)
Nucleosomal histones are barriers to the DNA repair process particularly at DNA double-strand breaks (DSBs). However, the molecular mechanism by which these histone barriers are removed from the sites of DNA damage remains elusive. Here, we have generated a single
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