17-650
ChIPAb+ Ubiquityl-Histone H2B - ChIP Validated Antibody and Primer Set
clone 56, from mouse, purified by using protein G
Synonym(s):
H2BUb, Histone H2B (ubiquityl), H2B histone family, member L, H2B.1 A, histone 1, H2bc, histone cluster 1, H2bc
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About This Item
UNSPSC Code:
12352203
eCl@ss:
32160702
NACRES:
NA.32
Recommended Products
biological source
mouse
Quality Level
clone
56, monoclonal
purified by
using protein G
species reactivity
mouse, rat, vertebrates, human, plant
manufacturer/tradename
ChIPAb+
Upstate®
technique(s)
ChIP: suitable
immunofluorescence: suitable
immunoprecipitation (IP): suitable
western blot: suitable
isotype
IgG2aκ
NCBI accession no.
UniProt accession no.
shipped in
dry ice
General description
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+ Ubiquityl-Histone H2B, (clone 56) set includes the Ubiquityl-Histone H2B, (clone 56) antibody, a negative control antibody (purified Mouse IgG), and qPCR primers which amplify a 213 bp region within the coding region of the human GAPDH gene.
The ubiquityl-Histone H2B and negative control antibodies are supplied in a scalable "per ChIP" reaction size and can be used to functionally validate the precipitation of Ubiquityl-Histone H2B associated chromatin.
The ChIPAb+ Ubiquityl-Histone H2B, (clone 56) set includes the Ubiquityl-Histone H2B, (clone 56) antibody, a negative control antibody (purified Mouse IgG), and qPCR primers which amplify a 213 bp region within the coding region of the human GAPDH gene.
The ubiquityl-Histone H2B and negative control antibodies are supplied in a scalable "per ChIP" reaction size and can be used to functionally validate the precipitation of Ubiquityl-Histone H2B associated chromatin.
Specificity
Not tested in other species. The immunogen sequence is identical in a wide range of animal and plant species, so broad cross-reactivity is expected.
Recognizes histone H2B ubiquitinated on lysine 120 but not free ubiquitin or histone H2B in its unmodified state.
Immunogen
Epitope: Lys120 ubiquitination site
Immunogen used was a branched peptide corresponding to the ubiquitination site on human histone H2B.
Application
Chromatin Immunoprecipitation:
Sonicated chromatin prepared from HeLa S3 cells (1 X 106 cell equivalents per IP) was subjected to chromatin immunoprecipitation using 1 μg of either a normal mouse IgG or Anti-Ubiquityl-Histone H2B antibody and the Magna ChIP G Kit (Cat. # 17-611).
Successful immunoprecipitation of ubiquityl-Histone H2B associated DNA fragments was verified by qPCR using GAPDH promoter (negative) and GAPDH coding (positive) 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. # 17-409) or EZ-ChIP (Cat. # 17-371) protocol for experimental details.
Western Blot Analysis:
HeLa cell lysate was resolved by electrophoresis, transferred to PVDF and probed with anti-Ubiquityl-histone H2B at a dilution of 1:4000 (Please see figures).
Sonicated chromatin prepared from HeLa S3 cells (1 X 106 cell equivalents per IP) was subjected to chromatin immunoprecipitation using 1 μg of either a normal mouse IgG or Anti-Ubiquityl-Histone H2B antibody and the Magna ChIP G Kit (Cat. # 17-611).
Successful immunoprecipitation of ubiquityl-Histone H2B associated DNA fragments was verified by qPCR using GAPDH promoter (negative) and GAPDH coding (positive) 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. # 17-409) or EZ-ChIP (Cat. # 17-371) protocol for experimental details.
Western Blot Analysis:
HeLa cell lysate was resolved by electrophoresis, transferred to PVDF and probed with anti-Ubiquityl-histone H2B at a dilution of 1:4000 (Please see figures).
Research Category
Epigenetics & Nuclear Function
Epigenetics & Nuclear Function
Research Sub Category
Chromatin Biology
Histones
Chromatin Biology
Histones
This ChIPAb+ Ubiquityl-Histone H2B -ChIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.
Packaging
25 assays per set. Recommended use: ~1 μg antibody per chromatin immunoprecipitation (dependent upon biological context).
Quality
Chromatin Immunoprecipitation:
Sonicated chromatin prepared from HeLa S3 cells (1X 106 cell equivalents per IP) were subjected to chromatin immuno-precipitation using 1 µg of either a normal mouse IgG or Anti-ubiquityl-Histone H2B, clone 56 antibody and the Magna ChIP® G Kit (Cat. # 17-611). Successful immunoprecipitation of ubiquityl-Histone H2B-associated DNA fragments was verified by qPCR using Control Primers.
Please refer to the EZ-Magna ChIP G (Cat. # 17-409) or EZ-ChIP (Cat. # 17-371) protocol for experimental details.
Sonicated chromatin prepared from HeLa S3 cells (1X 106 cell equivalents per IP) were subjected to chromatin immuno-precipitation using 1 µg of either a normal mouse IgG or Anti-ubiquityl-Histone H2B, clone 56 antibody and the Magna ChIP® G Kit (Cat. # 17-611). Successful immunoprecipitation of ubiquityl-Histone H2B-associated DNA fragments was verified by qPCR using Control Primers.
Please refer to the EZ-Magna ChIP G (Cat. # 17-409) or EZ-ChIP (Cat. # 17-371) protocol for experimental details.
Target description
~25 kDa
Physical form
Anti-ubiquityl-Histone H2B, clone 56 (mouse monoclonal IgG). One vial containing 25 μg of protein G purified antibody in 25 μg in 50 μL PBS containing 0.05% sodium azide. Store at -20°C.
Normal Mouse IgG. One vial containing 25 μg purified mouse IgG in 25 μL storage buffer containing 0.1% sodium azide. Store at -20°C.
ChIP Primers GAPDH Coding Region. One vial containing 75 μL of 5 μM of each primer specific for the coding region of human GAPDH. Store at -20°C.
FOR: GGC TCC CAC CTT TCT CAT CC
REV: GGC CAT CCA CAG TCT TCT GG
Normal Mouse IgG. One vial containing 25 μg purified mouse IgG in 25 μL storage buffer containing 0.1% sodium azide. Store at -20°C.
ChIP Primers GAPDH Coding Region. One vial containing 75 μL of 5 μM of each primer specific for the coding region of human GAPDH. Store at -20°C.
FOR: GGC TCC CAC CTT TCT CAT CC
REV: GGC CAT CCA CAG TCT TCT GG
Storage and Stability
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.
Analysis Note
Control
Includes negative control mouse IgG antibody and primers specific for human GAPDH coding region.
Includes negative control mouse IgG antibody and primers specific for human GAPDH coding region.
Legal Information
MAGNA CHIP is a registered trademark of Merck KGaA, Darmstadt, Germany
UPSTATE is a registered trademark of Merck KGaA, Darmstadt, Germany
Disclaimer
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.
Storage Class
10 - Combustible liquids
Certificates of Analysis (COA)
Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.
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