Anti-Histone H3 (citrulline R2 + R8 + R17) antibody - ChIP Grade (ab5103)
- Product nameAnti-Histone H3 (citrulline R2 + R8 + R17) antibody - ChIP GradeSee all Histone H3 primary antibodies ...
- DescriptionRabbit polyclonal to Histone H3 (citrulline R2 + R8 + R17) - ChIP Grade
- Specificityab5103 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 applicationsICC/IF, PepArr, IHC-Fr, ChIP/Chip, WB, ChIP, IHC-P more details
- Species reactivityReacts with: Mouse, Rat, Rabbit, Cow, Human, Monkey
Predicted to work with: a wide range of other species
Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human Histone H3, at Cit2, 8 and 17.
(Peptide available as ab32876.)
- Positive control
- This antibody gave a positive signal in HL60 Whole Cell Lysate - DMSO and Calcium Lonophore treated. In WB ab5103 only recognizes human or bovine histone H3 when PADI4 and calcium are added.
- Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
- Storage bufferPreservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4
- Concentration information loading...
- PurityImmunogen affinity purified
- Clonality Polyclonal
- Research Areas
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.
|ICC/IF||ICC/IF: Use at an assay dependent concentration. PubMed: 20733033|
|PepArr||PepArr: Use a concentration of 0.2 - 0.02 µg/ml.|
|IHC-Fr||IHC-Fr: Use at an assay dependent concentration.|
|ChIP/Chip||ChIP/Chip: Use at an assay dependent concentration.|
|WB||WB: Use a concentration of 1 µg/ml. Detects a band of approximately 17 kDa (predicted molecular weight: 15 kDa).Can be blocked with Human Histone H3 (citrulline 2 + 8 + 17) peptide (ab32876). Abcam recommends using 3-5% milk as the blocking agent|
|ChIP||ChIP: Use at an assay dependent concentration.|
|IHC-P||IHC-P: 1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.|
- RelevanceHistones are basic nuclear proteins that are responsible for the nucleosome structure of the chromosomal fiber in eukaryotes. Nucleosomes consist of approximately 146 bp of DNA wrapped around a histone octamer composed of pairs of each of the four core histones (H2A, H2B, H3, and H4). The chromatin fibre is further compacted through the interaction of a linker histone, H1, with the DNA between the nucleosomes to form higher order chromatin structures. Methylation of arginine residues within histone H3 has been linked to active transcription. However, the mechanism of demethylation of histones has remained elusive. Recently a process termed deimination has been described that converts histone arginine to citrulline and antagonizes arginine methylation (Cuthbert et al., 2004).
- Cellular localizationNuclear
- Entrez Gene: 326601 Cow
- Entrez Gene: 10338 Human
- Entrez Gene: 3020 Human
- Entrez Gene: 3021 Human
- Entrez Gene: 15079 Mouse
- Entrez Gene: 15080 Mouse
- Entrez Gene: 117056 Rat
- Entrez Gene: 289314 Rat
- Omim: 601128 Human
- SwissProt: P16105 Cow
- SwissProt: P16106 Cow
- SwissProt: P06351 Human
- SwissProt: P16106 Human
- SwissProt: Q16695 Human
- SwissProt: Q93081 Human
- SwissProt: A2A852 Mouse
- SwissProt: P16105 Mouse
- SwissProt: P84244 Mouse
- SwissProt: B0BMY8 Rat
- SwissProt: P06351 Rat
- SwissProt: P84245 Rat
- Unigene: 181307 Human
- Cit Histone H3 antibody
- Citrullated antibody
- citrullin antibody
- Citrullinated Histone H3 antibody
- citrulline antibody
- H3 3 like sequence MH921 antibody
- H3 3A antibody
- H3 histone antibody
- H3 histone family member E pseudogene antibody
- H3F3 antibody
- HIST3H3 antibody
- Histone H3 3 pseudogene antibody
- Histone H3 Citrullated antibody
Anti-Histone H3 (citrulline R2 + R8 + R17) antibody - ChIP Grade images
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
All lanes : Anti-Histone H3 (citrulline R2 + R8 + R17) antibody - ChIP Grade (ab5103) at 1 µg/ml
Lane 1 : HL60 (Human Caucasian promyelocytic leukaemia) DMSO and Calcium Lonophore treated Whole Cell Lysate (ab80919) with with 5% BSA
Lane 2 : HL60 (Human Caucasian promyelocytic leukaemia) DMSO and Calcium Lonophore treated Whole Cell Lysate (ab80919) with with 5% milk
Lane 3 : HL60 (Human Caucasian promyelocytic leukaemia) DMSO and Calcium Lonophore treated Whole Cell Lysate (ab80919) with with 3% milk
Lysates/proteins at 10 µg per lane.
Goat polyclonal Secondary Antibody to 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 seconds
Abcam recommends using milk as the blocking agent. Abcam welcomes customer feedback and would appreciate any comments regarding this product and the data presented above .
Chromatin immunoprecipitation using ab5103 on the pS2 promoter. Times are after stimulation by estrogen (Ul).
ICC/IF image of ab5103 stained human HeLa cells. The cells were PFA fixed (10 min), permabilised in TBS-T (20 min) and incubated with the antibody (ab5103, 1µg/ml) for 1h at room temperature. 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). DAPI was used to stain the cell nuclei (blue).
ab5103 staining Histone H3 (citrulline 2 + 8 + 17) in Mouse bone marrow cells by Immunocytochemistry/ Immunoflourescence. Cells were fixed in formaldehyde and permeabilized in 0.1% Triton X-100 prior to blocking in 5% Goat serum for 2 hours at 25°C. The primary antibody was diluted 1/250 in PBS and incubated with the sample for 12 hours at 4°C. The secondary antibody was Alexa Fluor® 488-conjugated Goat anti-Rabbit polyclonal, diluted 1/500.
Nuclei were counterstained blue with DAPI.
Rabbit polyclonal to Histone H3 (citrulline 2 + 8 + 17) used at 1/2000 dilution, after blocking with TBST 5% BSA. Purified histones run out with approximately 250 ng of each histone.
Lanes 1-3 contain Histone H3 (250 ng per lane)
Lane 1: PADI4 + Calcium
Lane 2: H3 + PADI4
Lane 3: H3 + PADI4 + Calcium
Lanes 4-5 contain bulk histones (250 ng per lane)
Lane 4: PADI4
Lane 5: PADI4 + Calcium
Lane 6: MCF7 cell extract
Lane 7: MCF7 cell extract (HA-PADI4)
Secondary antibody : anti-rabbit HRP from Sigma.
In WB ab5103 only recognizes human or bovine histone H3 when PADI4 and calcium are added.
References for Anti-Histone H3 (citrulline R2 + R8 + R17) antibody - ChIP Grade (ab5103)
This product has been referenced in:
- Neeli I & Radic M Opposition between PKC isoforms regulates histone deimination and neutrophil extracellular chromatin release. Front Immunol 4:38 (2013). Human . Read more (PubMed: 23430963) »
- Barrientos L et al. An improved strategy to recover large fragments of functional human neutrophil extracellular traps. Front Immunol 4:166 (2013). Read more (PubMed: 23805143) »