Recombinant Anti-Histone H2B (crotonyl K5) antibody [EPR17483] (ab177396)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR17483] to Histone H2B (crotonyl K5)
- Suitable for: PepArr, IHC-P, WB, ICC/IF
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-Histone H2B (crotonyl K5) antibody [EPR17483]
See all Histone H2B primary antibodies -
Description
Rabbit monoclonal [EPR17483] to Histone H2B (crotonyl K5) -
Host species
Rabbit -
Tested applications
Suitable for: PepArr, IHC-P, WB, ICC/IFmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: HeLa and NIH3T3 cell lysates;. IHC: Human colon, mouse colon, rat liver tissues. ICC/IF: HeLa cells.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR17483 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Isotype control
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab177396 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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PepArr |
Use at an assay dependent concentration.
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IHC-P |
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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WB |
1/500. Detects a band of approximately 14 kDa (predicted molecular weight: 14 kDa).
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ICC/IF |
1/250.
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Notes |
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PepArr
Use at an assay dependent concentration. |
IHC-P
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
WB
1/500. Detects a band of approximately 14 kDa (predicted molecular weight: 14 kDa). |
ICC/IF
1/250. |
Target
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Relevance
Core component of nucleosome. Nucleosomes wrap and compact DNA into chromatin, limiting DNA accessibility to the cellular machineries which require DNA as a template. Histones thereby play a central role in transcription regulation, DNA repair, DNA replication and chromosomal stability. DNA accessibility is regulated via a complex set of post-translational modifications of histones, also called histone code, and nucleosome remodeling. Subunit structure The nucleosome is a histone octamer containing two molecules each of H2A, H2B, H3 and H4 assembled in one H3-H4 heterotetramer and two H2A-H2B heterodimers. The octamer wraps approximately 147 bp of DNA. Post-translational modification Monoubiquitination at Lys-35 (H2BK34Ub) by the MSL1/MSL2 dimer is required for histone H3 'Lys-4' (H3K4me) and 'Lys-79' (H3K79me) methylation and transcription activation at specific gene loci, such as HOXA9 and MEIS1 loci. Similarly, monoubiquitination at Lys-121 (H2BK120Ub) by the RNF20/40 complex gives a specific tag for epigenetic transcriptional activation and is also prerequisite for histone H3 'Lys-4' and 'Lys-79' methylation. It also functions cooperatively with the FACT dimer to stimulate elongation by RNA polymerase II. H2BK120Ub also acts as a regulator of mRNA splicing: deubiquitination by USP49 is required for efficient cotranscriptional splicing of a large set of exons. Phosphorylation at Ser-37 (H2BS36ph) by AMPK in response to stress promotes transcription. Phosphorylated on Ser-15 (H2BS14ph) by STK4/MST1 during apoptosis; which facilitates apoptotic chromatin condensation. Also phosphorylated on Ser-15 in response to DNA double strand breaks (DSBs), and in correlation with somatic hypermutation and immunoglobulin class-switch recombination. GlcNAcylation at Ser-113 promotes monoubiquitination of Lys-121. It fluctuates in response to extracellular glucose, and associates with transcribed genes. Crotonylation (Kcr) is specifically present in male germ cells and marks testis-specific genes in post-meiotic cells, including X-linked genes that escape sex chromosome inactivation in haploid cells. Crotonylation marks active promoters and enhancers and confers resistance to transcriptional repressors. It is also associated with post-meiotically activated genes on autosomes. -
Cellular localization
Nuclear -
Database links
- Entrez Gene: 337874 Human
- Entrez Gene: 54145 Human
- Entrez Gene: 8349 Human
- Entrez Gene: 104021 Mouse
- Entrez Gene: 64647 Rat
- SwissProt: O60814 Human
- SwissProt: P06899 Human
- SwissProt: P23527 Human
see all -
Alternative names
- GL105 antibody
- H2B GL105 antibody
- H2B histone family member O antibody
see all
Images
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Anti-Histone H2B (crotonyl K5) antibody [EPR17483] (ab177396) at 1/1000 dilution + HeLa (Human epithelial cells from cervix adenocarcinoma) cell lysate at 20 µg
Secondary
Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution
Predicted band size: 14 kDa
Observed band size: 14 kDa
Exposure time: 3 minutesBlocking/Dilution buffer: 5% NFDM/TBST.
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Anti-Histone H2B (crotonyl K5) antibody [EPR17483] (ab177396) at 1/500 dilution + NIH/3T3 (Mouse embyro fibroblast cells) cell lysate at 10 µg
Secondary
Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution
Predicted band size: 14 kDa
Observed band size: 14 kDa
Exposure time: 3 minutesBlocking/Dilution buffer: 5% NFDM/TBST.
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ab177396 was tested in Peptide Array against 501 different modified and unmodified histone peptides; each peptide is printed on the array at six concentrations (each in triplicate).
Circle area represents affinity between the antibody and a peptide: all antigen-containing peptides are displayed as red circles, all other peptides as blue circles. The affinity is calculated as area under curve when antibody binding values are plotted against the corresponding peptide concentration. Each circle area is normalized to the peptide with the strongest affinity.
The complete dataset, including full list of all peptides and information on the position of each peptide in the diagram, can be downloaded here. -
Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling Histone H2B (crotonyl K5) using ab177396 at 1/500 dilution. A Goat Anti-Rabbit IgG H&L (HRP) (ab97051) was used as secondary at 1/500 dilution. Counter stained with Hematoxylin.
Inset image: negative control obtained using PBS instead of ab177396.
Note: Nucleus staining on glandular epithelium of Human colon tissue was observed.Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunohistochemical analysis of paraffin-embedded Mouse colon tissue labeling Histone H2B (crotonyl K5) using ab177396 at 1/500 dilution. A Goat Anti-Rabbit IgG H&L (HRP) (ab97051) was used as secondary at 1/500 dilution. Counter stained with Hematoxylin.
Inset image: negative control obtained using PBS instead of ab177396.
Note: Nucleus staining on glandular epithelium of Mouse colon tissue was observed.Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunohistochemical analysis of paraffin-embedded Rat liver tissue labeling Histone H2B (crotonyl K5) using ab177396 at 1/500 dilution. A Goat Anti-Rabbit IgG H&L (HRP) (ab97051) was used as secondary at 1/500 dilution. Counter stained with Hematoxylin.
Inset image: negative control obtained using PBS instead of ab177396.
Note: Nucleus staining on Rat liver tissue was observed.Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling Histone H2B (crotonyl K5) with ab177396 at 1/250 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green). Confocal image showing nuclear staining on HeLa cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).
The negative controls are as follows:
1. ab177396 at 1/250 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
2. ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500 dilution.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (0)
ab177396 has not yet been referenced specifically in any publications.