Recombinant Anti-Histone H2B (crotonyl K12) antibody [EPR17593] (ab177420)
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Overview
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Product name
Anti-Histone H2B (crotonyl K12) antibody [EPR17593]
See all Histone H2B primary antibodies -
Description
Rabbit monoclonal [EPR17593] to Histone H2B (crotonyl K12) -
Host species
Rabbit -
Tested applications
Suitable for: PepArr, IHC-P, WB, ICC/IFmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide within Human Histone H2B aa 1-100 (crotonyl K12). The exact sequence is proprietary.
Database link: Q16778 -
Positive control
- WB: NIH/3T3 and HeLa whole cell lysates. IHC-P: Human, mouse and rat colon tissues. ICC/IF: HeLa cells.
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General notes
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.
This product is a recombinant rabbit monoclonal antibody.
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, 0.05% BSA -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR17593 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Recombinant Protein
Applications
Our Abpromise guarantee covers the use of ab177420 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 |
|---|---|---|
| PepArr | Use at an assay dependent concentration. | |
| IHC-P | 1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. | |
| WB | 1/1000. Detects a band of approximately 14 kDa (predicted molecular weight: 14 kDa). | |
| ICC/IF | 1/100. |
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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![Western blot - Anti-Histone H2B (crotonyl K12) antibody [EPR17593] (ab177420)](https://www.abcam.com/ps/products/177/ab177420/Images/ab177420-233674-anti-histone-h2b-crotonyl-k12-antibody-epr17593-western-blot.jpg)
Western blot - Anti-Histone H2B (crotonyl K12) antibody [EPR17593] (ab177420)All lanes : Anti-Histone H2B (crotonyl K12) antibody [EPR17593] (ab177420) at 1/1000 dilution
Lane 1 : NIH/3T3 whole cell lysates
Lane 2 : HeLa whole cell lysates
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution
Predicted band size: 14 kDa
Observed band size: 14 kDaBlocking/Diluting buffer: 5% NFDM/TBST.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H2B (crotonyl K12) antibody [EPR17593] (ab177420)](https://www.abcam.com/ps/products/177/ab177420/Images/ab177420-233678-anti-histone-h2b-crotonyl-k12-antibody-epr17593-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H2B (crotonyl K12) antibody [EPR17593] (ab177420)Immunohistochemical analysis of paraffin-embedded rat colon tissue labeling Histone H2B (crotonyl K12) with ab177420 at 1/1000 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Nucleus staining on glandular epithelium of rat colon tissue is observed. Counter stained with Hematoxylin.
Negative control: Using PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution.Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H2B (crotonyl K12) antibody [EPR17593] (ab177420)](https://www.abcam.com/ps/products/177/ab177420/Images/ab177420-233677-anti-histone-h2b-crotonyl-k12-antibody-epr17593-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H2B (crotonyl K12) antibody [EPR17593] (ab177420)Immunohistochemical analysis of paraffin-embedded mouse colon tissue labeling Histone H2B (crotonyl K12) with ab177420 at 1/1000 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Nucleus staining on glandular epithelium of mouse colon tissue is observed. Counter stained with Hematoxylin.
Negative control: Using PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution.Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H2B (crotonyl K12) antibody [EPR17593] (ab177420)](https://www.abcam.com/ps/products/177/ab177420/Images/ab177420-233676-anti-histone-h2b-crotonyl-k12-antibody-epr17593-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H2B (crotonyl K12) antibody [EPR17593] (ab177420)Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling Histone H2B (crotonyl K12) with ab177420 at 1/1000 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Nucleus staining on glandular epithelium of Human colon tissue is observed. Counter stained with Hematoxylin.
Negative control: Using PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution.Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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![Immunocytochemistry/ Immunofluorescence - Anti-Histone H2B (crotonyl K12) antibody [EPR17593] (ab177420)](https://www.abcam.com/ps/products/177/ab177420/Images/ab177420-244258-anti-histone-h2b-crotonyl-k12-antibody-epr17593-immunofluorescence.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-Histone H2B (crotonyl K12) antibody [EPR17593] (ab177420)Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa cells labeling Histone H2B (crotonyl K12) with ab177420 at 1/100 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/400 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 1/500 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).
The negative controls are as follows:
-ve control 1: ab177420 at 1/100 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
-ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/400 dilution. -
![Peptide Array - Anti-Histone H2B (crotonyl K12) antibody [EPR17593] (ab177420)](https://www.abcam.com/ps/products/177/ab177420/Images/ab177420-233675-anti-histone-h2b-crotonyl-k12-antibody-epr17593-peptide-array.jpg)
Peptide Array - Anti-Histone H2B (crotonyl K12) antibody [EPR17593] (ab177420)ab177420 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.
Protocols
Datasheets and documents
References
ab177420 has not yet been referenced specifically in any publications.
