Anti-Histone H3 (citrulline R2 + R8 + R17) antibody (ab5103)
Key features and details
- Rabbit polyclonal to Histone H3 (citrulline R2 + R8 + R17)
- Suitable for: ICC/IF, PepArr, IP, WB
- Reacts with: Mouse, Rat, Human, Recombinant fragment
- Isotype: IgG
Get better batch-to-batch reproducibility with a recombinant antibody
- Research with confidence – consistent and reproducible results with every batch
- Long-term and scalable supply – powered by recombinant technology for fast production
- Success from the first experiment – confirmed specificity through extensive validation
- Ethical standards compliant – production is animal-free
Overview
-
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
Suitable for: ICC/IF, PepArr, IP, WBmore details -
Species reactivity
Reacts with: Mouse, Rat, Human, Recombinant fragment
Predicted to work with: Rabbit, Cow, Monkey, a wide range of other species
-
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
(Peptide available asab32876) -
Positive control
- WB: Mouse and rat brain tissue lysates. HL60 + DMSO, NIH/3T3 and PC12 whole cell lysates. ICC/IF: MCF7 cells. IP: HEK-293T (human embryonic kidney) transfected with PADI4 expression vector containing a GFP-tag
-
General notes
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
Properties
-
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
-
ChIP Related Products
-
Compatible Secondaries
-
Immunizing Peptide (Blocking)
-
Isotype control
-
Recombinant Protein
-
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.
| Application | Abreviews | Notes |
|---|---|---|
| ICC/IF | (8) |
Use a concentration of 1 µg/ml.
|
| PepArr |
Use a concentration of 0.2 - 0.02 µg/ml.
|
|
| IP |
Use at an assay dependent concentration.
|
|
| 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 |
|---|
|
ICC/IF
Use a concentration of 1 µg/ml. |
|
PepArr
Use a concentration of 0.2 - 0.02 µg/ml. |
|
IP
Use at an assay dependent concentration. |
|
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
-
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
-
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
-
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
-
Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (citrulline R2 + R8 + R17) antibody (ab5103)Immunocytochemical anaylsis of paraformaldehyde-fixed human HeLa cells, permeabilised in TBST (20 min) and incubated with ab5103 at 1µg/ml for 1h at RT. 1% BSA / 10% normal goat serum / 0.3M glycine was used to quench autofluorescence and block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). The nucleus was stained with DAPI (blue).
-
Western blot - Anti-Histone H3 (citrulline R2 + R8 + R17) antibody (ab5103)This western blot image is a comparison between ab5103 and the alternative recombinant multiclonal antibody ab281584.
Left side - Recombinant multiclonal to Histone H3 (citrulline R2 + R8 + R17) - ab281584
All lanes: Anti-Histone H3 (citrulline R2 + R8 + R17) antibody [RM1001] ab281584 at 1/1000 dilution.
Lane 1: Untreated HEK-293T (human embryonic kidney) transfected with PADI4 expression vector containing a myc-His-tag®, whole cell lysate
Lane 2: HEK-293T transfected with PADI4 expression vector containing a myc-His-tag® treated with 10 mM calcium chloride and 10 µM lonomycin for 4 hours, whole cell lysate
Lane 3: Untreated NIH/3T3 (mouse embryonic fibroblast) transfected with PADI4 expression vector containing a myc-His-tag®, whole cell lysate
Lane 4: NIH/3T3 transfected with PADI4 expression vector containing a myc-His-tag® treated with 10 mM calcium chloride and 10 µM lonomycin for 4 hours, whole cell lysateRight side: Anti-Histone H3 (citrulline R2 + R8 + R17) antibody (ab5103)
All lanes: Anti-Histone H3 (citrulline R2 + R8 + R17) antibody ab5103 at 1/1000 dilution.
Lane 1: Untreated HEK-293T (human embryonic kidney) transfected with PADI4 expression vector containing a myc-His-tag®, whole cell lysate
Lane 2: HEK-293T transfected with PADI4 expression vector containing a myc-His-tag® treated with 10 mM calcium chloride and 10 µM lonomycin for 4 hours, whole cell lysate
Lane 3: Untreated NIH/3T3 (mouse embryonic fibroblast) transfected with PADI4 expression vector containing a myc-His-tag®, whole cell lysate
Lane 4: NIH/3T3 transfected with PADI4 expression vector containing a myc-His-tag® treated with 10 mM calcium chloride and 10 µM lonomycin for 4 hours, whole cell lysateLysates/proteins at 20 µg per lane.
Secondary
All lanes: Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)Why choose a recombinant antibody?
Research with confidence – consistent and reproducible results with every batch
Long-term and scalable supply – powered by recombinant technology for fast production
Success from the first experiment – confirmed specificity through extensive validation
Ethical standards compliant – production is animal-free -
Immunoprecipitation - Anti-Histone H3 (citrulline R2 + R8 + R17) antibody (ab5103)This immunoprecipitation image is a comparison between ab5103 and the alternative recombinant multiclonal antibody ab281584.
Left side - Recombinant multiclonal to Histone H3 (citrulline R2 + R8 + R17) - ab281584
All lanes: Anti-Histone H3 (citrulline R2 + R8 + R17) antibody [RM1001] ab281584 at 1/1000 dilution.
Lane 1: HEK-293T (human embryonic kidney) transfected with PADI4 expression vector containing a GFP-tag treated with 10 mM calcium chloride and 10 μM lonomycin for 4 hours, whole cell lysate 10 μg.
Lane 2: ab281584 IP in HEK-293T transfected with PADI4 expression vector containing a GFP-tag treated with 10 mM calcium chloride and 10 μM lonomycin for 4 hours whole cell lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab281584 in HEK-293T transfected with PADI4 expression vector containing a GFP-tag treated with 10 mM calcium chloride and 10 μM lonomycin for 4 hours whole cell lysateRight side: Anti-Histone H3 (citrulline R2 + R8 + R17) antibody (ab5103)
All lanes: Anti-Histone H3 (citrulline R2 + R8 + R17) antibody ab5103 at 1/1000 dilution.
Lane 1: HEK-293T (human embryonic kidney) transfected with PADI4 expression vector containing a GFP-tag treated with 10 mM calcium chloride and 10 μM lonomycin for 4 hours, whole cell lysate 10 μg.
Lane 2: ab5103 IP in HEK-293T transfected with PADI4 expression vector containing a GFP-tag treated with 10 mM calcium chloride and 10 μM lonomycin for 4 hours whole cell lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab5103 in HEK-293T transfected with PADI4 expression vector containing a GFP-tag treated with 10 mM calcium chloride and 10 μM lonomycin for 4 hours whole cell lysateSecondary
All lanes: VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.
Predicted band size: 15kDaWhy choose a recombinant antibody?
Research with confidence – consistent and reproducible results with every batch
Long-term and scalable supply – powered by recombinant technology for fast production
Success from the first experiment – confirmed specificity through extensive validation
Ethical standards compliant – production is animal-free -
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
-
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.
-
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
-
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
Datasheets and documents
-
SDS download
-
Datasheet download
References (352)
ab5103 has been referenced in 352 publications.
- Chirivi RGS et al. Therapeutic ACPA inhibits NET formation: a potential therapy for neutrophil-mediated inflammatory diseases. Cell Mol Immunol 18:1528-1544 (2021). PubMed: 32203195
- Locke M & Longstaff C Extracellular Histones Inhibit Fibrinolysis through Noncovalent and Covalent Interactions with Fibrin. Thromb Haemost 121:464-476 (2021). PubMed: 33131044
- Takeuchi M et al. Neutrophils interact with cholangiocytes to cause cholestatic changes in alcoholic hepatitis. Gut 70:342-356 (2021). PubMed: 33214166
- Mondal S et al. Site-specific incorporation of citrulline into proteins in mammalian cells. Nat Commun 12:45 (2021). PubMed: 33398026
- Shi Y et al. Neutrophils can promote clotting via FXI and impact clot structure via neutrophil extracellular traps in a distinctive manner in vitro. Sci Rep 11:1718 (2021). PubMed: 33462294