Recombinant Anti-Histone H3 (citrulline R2 + R8 + R17) antibody [RM1001] (ab281584)
![Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (citrulline R2 + R8 + R17) antibody [RM1001] (ab281584)](https://www.abcam.com/ps/products/281/ab281584/Images/ab281584-488340-anti-histone-h3-citrulline-r2-r8-r17-antibody-rm1001-immunocytochemistry-293t-cells-human.jpg "Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (citrulline R2 + R8 + R17) antibody [RM1001] (ab281584)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit recombinant multiclonal [RM1001] to Histone H3 (citrulline R2 + R8 + R17)
- Suitable for: IP, PepArr, ICC/IF, Flow Cyt (Intra), WB, Dot blot
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
-
Product name
Anti-Histone H3 (citrulline R2 + R8 + R17) antibody [RM1001]
See all Histone H3 primary antibodies -
Description
Rabbit recombinant multiclonal [RM1001] to Histone H3 (citrulline R2 + R8 + R17) -
Host species
Rabbit -
Tested applications
Suitable for: IP, PepArr, ICC/IF, Flow Cyt (Intra), WB, Dot blotmore details
Unsuitable for: ChIP or IHC-P -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
This product was produced with the following immunogens:
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers. -
Positive control
- WB: HEK-293T, NIH/3T3 cells transfected with PADI4 expression vector containing a myc-His-tag® treated with 10 mM CaCl2 and 10 µM lonomycin for 4 h, C6 cells transfected with PADI4 expression vector containing a GFP-tag treated with 10 mM CaCl2 and 10 µM lonomycin for 2 h. IP: HEK-293T cells. ICC/IF: 293T cells. Flow Cyt (Intra): 293T cells transfected with myc-tagged PADI4 construct, then treated with 10mM CaCl2 and 10uM Ionomycin for 4h.
-
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.
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 long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Recombinant Multiclonal -
Clone number
RM1001 -
Isotype
IgG -
Research areas
Associated products
-
Compatible Secondaries
-
Isotype control
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab281584 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 |
|---|---|---|
| IP |
1/30.
|
|
| PepArr |
Use at an assay dependent concentration.
|
|
| ICC/IF |
1/2000.
|
|
| Flow Cyt (Intra) |
1/500.
|
|
| WB |
1/1000. Predicted molecular weight: 15 kDa.
|
|
| Dot blot |
1/1000.
|
| Notes |
|---|
|
IP
1/30. |
|
PepArr
Use at an assay dependent concentration. |
|
ICC/IF
1/2000. |
|
Flow Cyt (Intra)
1/500. |
|
WB
1/1000. Predicted molecular weight: 15 kDa. |
|
Dot blot
1/1000. |
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
-
Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (citrulline R2 + R8 + R17) antibody [RM1001] (ab281584)
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 100% methanol permeabilized 293T (human embryonic kidney epithelial cell) cells labelling Histone H3 (citrulline R2 + R8 + R17) with primary antibody anti-Histone H3 (citrulline R2 + R8 + R17) (ab281584) at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) (ab150080) secondary antibody at 1/1000 dilution. Confocal image showing nuclear staining on 293T cells transfected with a human PADI4 expression vector containing a GFP tag, then treated with 10 mM CaCl2 and 10 μM ionomycin for 2 hours. The nuclear counter stain is DAPI (blue).
-
![Western blot - Anti-Histone H3 (citrulline R2 + R8 + R17) antibody [RM1001] (ab281584)](https://www.abcam.com/ps/products/281/ab281584/Images/ab281584-459113-anti-histone-h3-citrulline-r2--r8--r17-antibody-oligo1-western-blot-glial.jpg)
Western blot - Anti-Histone H3 (citrulline R2 + R8 + R17) antibody [RM1001] (ab281584)All lanes : Anti-Histone H3 (citrulline R2 + R8 + R17) antibody [RM1001] (ab281584) at 1/1000 dilution
Lane 1 : Untreated C6 (rat glial tumor glial cell) transfected with PADI4 expression vector containing a GFP-tag, whole cell lysate
Lane 2 : C6 transfected with PADI4 expression vector containing a GFP-tag treated with 10 mM calcium chloride and 10 µM lonomycin for 2 hours, whole cell lysate
Lysates/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)
Predicted band size: 15 kDaBlocking and diluting buffer and concentration: 5% NFDM/TBST
Exposure time: 180 seconds
-
![Peptide Array - Anti-Histone H3 (citrulline R2 + R8 + R17) antibody [RM1001] (ab281584)](https://www.abcam.com/ps/products/281/ab281584/Images/ab281584-503810-anti-histone-h3-citrulline-r2-r8-r17-antibody-rm1001-peptide-array.png)
Peptide Array - Anti-Histone H3 (citrulline R2 + R8 + R17) antibody [RM1001] (ab281584)Peptide array analysis of ab281584 at (0.1ug/ml) followed by a Goat Anti-Rabbit IgG, (H+L), Fluo 647nm conjugated at 1:50,000 dilution.
All batches of ab281584 are 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 -
![Flow Cytometry (Intracellular) - Anti-Histone H3 (citrulline R2 + R8 + R17) antibody [RM1001] (ab281584)](https://www.abcam.com/ps/products/281/ab281584/Images/ab281584-488354-anti-histone-h3-citrulline-r2r8r17-antibody-rm1001-flowcytometry-human.jpg)
Flow Cytometry (Intracellular) - Anti-Histone H3 (citrulline R2 + R8 + R17) antibody [RM1001] (ab281584)Intracellular Flow Cytometry analysis of 293T (human embryonic kidney epithelial cell) transfected with myc-tagged PADI4 construct, then treated with 10mM CaCl2 and 10uM Ionomycin for 4h labeling Histone H3 (citrulline R2 + R8 + R17) with ab281584 at 1/500 dilution. Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as a secondary antibody.
Isotype control : Rabbit monoclonal IgG (ab172730)/left.
-
![Western blot - Anti-Histone H3 (citrulline R2 + R8 + R17) antibody [RM1001] (ab281584)](https://www.abcam.com/ps/products/281/ab281584/Images/ab281584-459112-anti-histone-h3-citrulline-r2--r8--r17-antibody-oligo1-western-blot-embryonic.jpg)
Western blot - Anti-Histone H3 (citrulline R2 + R8 + R17) antibody [RM1001] (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 lysate
Lysates/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)
Predicted band size: 15 kDaBlocking and diluting buffer and concentration: 5% NFDM/TBST
Exposure time: 48 seconds
-
![Dot Blot - Anti-Histone H3 (citrulline R2 + R8 + R17) antibody [RM1001] (ab281584)](https://www.abcam.com/ps/products/281/ab281584/Images/ab281584-2-anti-histone-h3-citrulline-r2--r8--r17-antibody-oligo1-dotblot-histone-h3.jpg)
Dot Blot - Anti-Histone H3 (citrulline R2 + R8 + R17) antibody [RM1001] (ab281584)Dot blot analysis of Histone H3 (citrulline R2 + R8 + R17) using 281584 at 1/1000 (0.478 ug/ml) followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100,000 dilution.
Lane 1: Histone H3 (citrulline R2) peptide (aa 1-21)
Lane 2: Histone H3 (citrulline R8) peptide (aa 1-21)
Lane 3: Histone H3 (citrulline R17) peptide (aa 1-21)
Lane 4: Histone H3 (citrulline R2/R8/R17) peptide (aa 1-21)
Lane 5: Unmodified histone H3 peptide (aa 1-21)
Exposure time: 3 minutes
Blocking and diluting buffer and concentration: 5% NFDM/TBST
-
![Immunoprecipitation - Anti-Histone H3 (citrulline R2 + R8 + R17) antibody [RM1001] (ab281584)](https://www.abcam.com/ps/products/281/ab281584/Images/ab281584-1-anti-histone-h3-citrulline-r2--r8--r17-antibody-oligo1-immunoprecipitation-hek293t.jpg)
Immunoprecipitation - Anti-Histone H3 (citrulline R2 + R8 + R17) antibody [RM1001] (ab281584)Histone H3 (citrulline R2 + R8 + R17) was immunoprecipitated from 0.35 mg 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 with 281584 at 1/30 dilution (2μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using 281584 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 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 lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 10 seconds -
![Anti-Histone H3 (citrulline R2 + R8 + R17) antibody [RM1001] (ab281584)](https://www.abcam.com/ps/products/281/ab281584/Images/ab281584-3-Benefits-of-Recombinant-antibodies.jpg)
Anti-Histone H3 (citrulline R2 + R8 + R17) antibody [RM1001] (ab281584)
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
-
SDS download
-
Datasheet download
References (0)
ab281584 has not yet been referenced specifically in any publications.