Anti-Histone H3 (citrulline R2 + R8 + R17) antibody (ab5103)
Key features and details
- Rabbit polyclonal to Histone H3 (citrulline R2 + R8 + R17)
- Suitable for: PepArr, ICC, WB
- Reacts with: Mouse, Rat, Human
- Isotype: IgG
Get better batch-to-batch reproducibility with a recombinant antibody
- Research with confidence – consistent and reproducible results with every batch
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- Success from the first experiment – confirmed specificity through extensive validation
- Ethical standards compliant – production is animal-free
Overview
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Product name
Anti-Histone H3 (citrulline R2 + R8 + R17) antibody
See all Histone H3 primary antibodies -
Description
Rabbit polyclonal to Histone H3 (citrulline R2 + R8 + R17) -
Host species
Rabbit -
Specificity
ab5103 detects a 17 kDa band in single lane Western Blot. Peptide inhibition in Western Blot hasn't been processed. Modification specificity is determined by Peptide Array. ab5103 binds strongly to Histone H3 citrulline 2 + 8 + 17 peptide. -
Tested Applications & Species
See all applications and species dataApplication Species ICC HumanWB MouseRatHuman
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Immunogen
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Positive control
- WB: Mouse and rat brain tissue lysates. HL60 + DMSO, NIH/3T3 and PC12 whole cell lysates. ICC: MCF7 cells.
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General notes
Immunizing Peptide (Blocking) - ab32876.
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 or -80°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS
Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help. -
Concentration information loading... -
Purity
Immunogen affinity purified -
Clonality
Polyclonal -
Isotype
IgG -
Research areas
Associated products
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ChIP Related Products
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Compatible Secondaries
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Immunizing Peptide (Blocking)
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Isotype control
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Recombinant Protein
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Related Products
- Anti-Histone H3 antibody [mAbcam 10799] - ChIP Grade (ab10799)
- Prestained Protein Ladder - Broad molecular weight (10-245 kDa) (ab116028)
- Anti-Histone H3 (di methyl K9) antibody [mAbcam 1220] - ChIP Grade (ab1220)
- Human Histone H3 (tri methyl K4) peptide (ab1342)
- Anti-Histone H3 (phospho S10) antibody [mAbcam 14955] (ab14955)
- Human Histone H3 (tri methyl K9) peptide (ab1773)
- Human Histone H3 (tri methyl K27) peptide (ab1782)
- Anti-Histone H3 (tri methyl K27) antibody [mAbcam 6002] - ChIP Grade (ab6002)
- Human Histone H3 (unmodified) peptide (ab7228)
- Human Histone H3 (di methyl K4) peptide (ab7768)
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab5103 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Tested applications are guaranteed to work and covered by our Abpromise guarantee.
Predicted to work for this combination of applications and species but not guaranteed.
Does not work for this combination of applications and species.
| Application | Species |
|---|---|
| ICC |
Human
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| WB |
Mouse
Rat
Human
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| All applications |
Rabbit
Cow
Monkey
a wide range of other species
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| Application | Abreviews | Notes |
|---|---|---|
| PepArr |
Use a concentration of 0.2 - 0.02 µg/ml.
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| ICC |
Use a concentration of 1 µg/ml.
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| WB | (9) |
Use a concentration of 1 µg/ml. Detects a band of approximately 17 kDa (predicted molecular weight: 15 kDa).
In our hands, when tested in western blot, this product typically gives a weaker signal in mouse and rat tissue lysates compared to mouse and rat cell lines. Abcam welcomes customer feedback and would appreciate any comments regarding this product and the data presented above. Abcam recommends using 3-5% milk as the blocking agent We recommend Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody. |
| Notes |
|---|
|
PepArr
Use a concentration of 0.2 - 0.02 µg/ml. |
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ICC
Use a concentration of 1 µg/ml. |
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WB
Use a concentration of 1 µg/ml. Detects a band of approximately 17 kDa (predicted molecular weight: 15 kDa). In our hands, when tested in western blot, this product typically gives a weaker signal in mouse and rat tissue lysates compared to mouse and rat cell lines. Abcam welcomes customer feedback and would appreciate any comments regarding this product and the data presented above. Abcam recommends using 3-5% milk as the blocking agent We recommend Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody. |
Target
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Function
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. -
Sequence similarities
Belongs to the histone H3 family. -
Developmental stage
Expressed during S phase, then expression strongly decreases as cell division slows down during the process of differentiation. -
Post-translational
modificationsAcetylation is generally linked to gene activation. Acetylation on Lys-10 (H3K9ac) impairs methylation at Arg-9 (H3R8me2s). Acetylation on Lys-19 (H3K18ac) and Lys-24 (H3K24ac) favors methylation at Arg-18 (H3R17me).
Citrullination at Arg-9 (H3R8ci) and/or Arg-18 (H3R17ci) by PADI4 impairs methylation and represses transcription.
Asymmetric dimethylation at Arg-18 (H3R17me2a) by CARM1 is linked to gene activation. Symmetric dimethylation at Arg-9 (H3R8me2s) by PRMT5 is linked to gene repression. Asymmetric dimethylation at Arg-3 (H3R2me2a) by PRMT6 is linked to gene repression and is mutually exclusive with H3 Lys-5 methylation (H3K4me2 and H3K4me3). H3R2me2a is present at the 3' of genes regardless of their transcription state and is enriched on inactive promoters, while it is absent on active promoters.
Methylation at Lys-5 (H3K4me), Lys-37 (H3K36me) and Lys-80 (H3K79me) are linked to gene activation. Methylation at Lys-5 (H3K4me) facilitates subsequent acetylation of H3 and H4. Methylation at Lys-80 (H3K79me) is associated with DNA double-strand break (DSB) responses and is a specific target for TP53BP1. Methylation at Lys-10 (H3K9me) and Lys-28 (H3K27me) are linked to gene repression. Methylation at Lys-10 (H3K9me) is a specific target for HP1 proteins (CBX1, CBX3 and CBX5) and prevents subsequent phosphorylation at Ser-11 (H3S10ph) and acetylation of H3 and H4. Methylation at Lys-5 (H3K4me) and Lys-80 (H3K79me) require preliminary monoubiquitination of H2B at 'Lys-120'. Methylation at Lys-10 (H3K9me) and Lys-28 (H3K27me) are enriched in inactive X chromosome chromatin.
Phosphorylated at Thr-4 (H3T3ph) by GSG2/haspin during prophase and dephosphorylated during anaphase. Phosphorylation at Ser-11 (H3S10ph) by AURKB is crucial for chromosome condensation and cell-cycle progression during mitosis and meiosis. In addition phosphorylation at Ser-11 (H3S10ph) by RPS6KA4 and RPS6KA5 is important during interphase because it enables the transcription of genes following external stimulation, like mitogens, stress, growth factors or UV irradiation and result in the activation of genes, such as c-fos and c-jun. Phosphorylation at Ser-11 (H3S10ph), which is linked to gene activation, prevents methylation at Lys-10 (H3K9me) but facilitates acetylation of H3 and H4. Phosphorylation at Ser-11 (H3S10ph) by AURKB mediates the dissociation of HP1 proteins (CBX1, CBX3 and CBX5) from heterochromatin. Phosphorylation at Ser-11 (H3S10ph) is also an essential regulatory mechanism for neoplastic cell transformation. Phosphorylated at Ser-29 (H3S28ph) by MLTK isoform 1, RPS6KA5 or AURKB during mitosis or upon ultraviolet B irradiation. Phosphorylation at Thr-7 (H3T6ph) by PRKCBB is a specific tag for epigenetic transcriptional activation that prevents demethylation of Lys-5 (H3K4me) by LSD1/KDM1A. At centromeres, specifically phosphorylated at Thr-12 (H3T11ph) from prophase to early anaphase, by DAPK3 and PKN1. Phosphorylation at Thr-12 (H3T11ph) by PKN1 is a specific tag for epigenetic transcriptional activation that promotes demethylation of Lys-10 (H3K9me) by KDM4C/JMJD2C. Phosphorylation at Tyr-42 (H3Y41ph) by JAK2 promotes exclusion of CBX5 (HP1 alpha) from chromatin.
Monoubiquitinated by RAG1 in lymphoid cells, monoubiquitination is required for V(D)J recombination (By similarity). Ubiquitinated by the CUL4-DDB-RBX1 complex in response to ultraviolet irradiation. This may weaken the interaction between histones and DNA and facilitate DNA accessibility to repair proteins. -
Cellular localization
Nucleus. Chromosome. - Information by UniProt
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Database links
- Entrez Gene: 8350 Human
- Entrez Gene: 8351 Human
- Entrez Gene: 8352 Human
- Entrez Gene: 8353 Human
- Entrez Gene: 8354 Human
- Entrez Gene: 8355 Human
- Entrez Gene: 8356 Human
- Entrez Gene: 8357 Human
see all -
Alternative names
- H3 histone family member E pseudogene antibody
- H3 histone family, member A antibody
- H3/A antibody
see all
Images
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Western blot - Anti-Histone H3 (citrulline R2 + R8 + R17) antibody (ab5103)All lanes : Anti-Histone H3 (citrulline R2 + R8 + R17) antibody (ab5103) at 1 µg/ml
Lane 1 : Mouse brain tissue lysate
Lane 2 : Rat brain tissue lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/50000 dilution
Predicted band size: 15 kDa
Observed band size: 17 kDa why is the actual band size different from the predicted?
Exposure time: 1 minuteBlocking buffer: 2% BSA
Gel type: MES
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Immunocytochemistry - Anti-Histone H3 (citrulline R2 + R8 + R17) antibody (ab5103)Immunocytochemistry analysis of ab5103 staining Histone H3 (citrulline R2 + R8 + R17) in MCF7 cells. The cells were fixed with 4% paraformaldehyde (10 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab5103 at 1µg/mL and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488), pre-adsorbed at 1/1000 dilution (shown in green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 594), pre-adsorbed at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue). Also suitable in cells fixed with 100% methanol (5 min).
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Western blot - Anti-Histone H3 (citrulline R2 + R8 + R17) antibody (ab5103)All lanes : Anti-Histone H3 (citrulline R2 + R8 + R17) antibody (ab5103) at 1 µg/ml
Lane 1 : NIH/3T3 (Mouse embryo fibroblast cell line) nuclear lysate
Lane 2 : PC12 (Rat adrenal pheochromocytoma cell line) nuclear lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/50000 dilution
Predicted band size: 15 kDa
Observed band size: 17 kDa why is the actual band size different from the predicted?
Exposure time: 1 minuteBlocking buffer: 2% BSA
Gel type: MES
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Western blot - Anti-Histone H3 (citrulline R2 + R8 + R17) antibody (ab5103)All lanes : Anti-Histone H3 (citrulline R2 + R8 + R17) antibody (ab5103) at 0.2 µg/ml
Lane 1 : HL60 whole cell lysate (negative control)
Lane 2 : HL60 whole cell lysate + DMSO (solvent control)
Lane 3 : HL60 whole cell lysate + DMSO + Calcium Ionophore (positive control)
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat anti Rabbit IR680 at 1/10000 dilution
Performed under reducing conditions.
Predicted band size: 15 kDa
Observed band size: 17 kDa why is the actual band size different from the predicted?Loading Control: GAPDH
This blot was produced using a 4-12% Bis-tris gel under the MES buffer system. The gel was run at 200V for 35 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using Licor blocking buffer before being incubated with ab5103 overnight at 4°C. Antibody binding was detected using Goat anti Rabbit IR680 secondary at a 1:10,000 dilution for 1hr at room temperature and then imaged using the Licor Odyssey CLx.
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Peptide Array - Anti-Histone H3 (citrulline R2 + R8 + R17) antibody (ab5103)All batches of ab5103 are tested in Peptide Array against peptides to different Histone H3 modifications. Six dilutions of each peptide are printed on to the Peptide Array in triplicate and results are averaged before being plotted on to a graph. Results show strong binding to Histone H3 - citrulline 2 + 8 + 17 peptide (ab32876), indicating that this antibody specifically recognises the Histone H3 - citrulline 2 + 8 + 17 modifications.
ab32876 - Histone H3 - citrulline 2 + 8 + 17
ab17566 - Histone H3 - unmodified
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Western blot - Anti-Histone H3 (citrulline R2 + R8 + R17) antibody (ab5103)All lanes : Anti-Histone H3 (citrulline R2 + R8 + R17) antibody (ab5103) at 1 µg/ml
Lane 1 : HL60 (Human Caucasian promyelocytic leukaemia) DMSO and Calcium Ionophore treated Whole Cell Lysate with with 5% BSA
Lane 2 : HL60 (Human Caucasian promyelocytic leukaemia) DMSO and Calcium Ionophore treated Whole Cell Lysate with with 5% milk
Lane 3 : HL60 (Human Caucasian promyelocytic leukaemia) DMSO and Calcium Ionophore treated Whole Cell Lysate with with 3% milk
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 15 kDa
Observed band size: 17 kDa why is the actual band size different from the predicted?
Exposure time: 30 secondsAbcam recommends using milk as the blocking agent. Abcam welcomes customer feedback and would appreciate any comments regarding this product and the data presented above .
Blots were developled with Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody
Protocols
References (250)
ab5103 has been referenced in 250 publications.
- Jackson-Jones LH et al. Stromal Cells Covering Omental Fat-Associated Lymphoid Clusters Trigger Formation of Neutrophil Aggregates to Capture Peritoneal Contaminants. Immunity 52:700-715.e6 (2020). PubMed: 32294409
- Alhede M et al. The origin of extracellular DNA in bacterial biofilm infections in vivo. Pathog Dis 78:N/A (2020). PubMed: 32196074
- Song H et al. HNF4A-AS1/hnRNPU/CTCF axis as a therapeutic target for aerobic glycolysis and neuroblastoma progression. J Hematol Oncol 13:24 (2020). PubMed: 32216806
- de Bont CM et al. Neutrophil proteases degrade autoepitopes of NET-associated proteins. Clin Exp Immunol 199:1-8 (2020). PubMed: 31661552
- Chatterjee V et al. Endothelial microvesicles carrying Src-rich cargo impair adherens junction integrity and cytoskeleton homeostasis. Cardiovasc Res 116:1525-1538 (2020). PubMed: 31504252
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