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- Histone H3 (citrulline 2 + 8 + 17) antibody [CitH3] - ChIP Grade (ab5103)
![- Histone H3 (citrulline 2 + 8 + 17) antibody [CitH3] - ChIP Grade (ab5103)](/ps/datasheet/images/ab5103_1.jpg)
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Immunocytochemistry/ Immunofluorescence - Histone H3 (citrulline 2 + 8 + 17) antibody [CitH3] - ChIP Grade (ab5103)
![Immunocytochemistry/ Immunofluorescence - Histone H3 (citrulline 2 + 8 + 17) antibody [CitH3] - ChIP Grade (ab5103)](/ps/datasheet/images/ab5103-IF-Im1.jpg)
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Western blot - Histone H3 (citrulline 2 + 8 + 17) antibody [CitH3] - ChIP Grade (ab5103)
![Western blot - Histone H3 (citrulline 2 + 8 + 17) antibody [CitH3] - ChIP Grade (ab5103)](/ps/datasheet/images/5/ab5103/Histone-H3-Primary-antibodies-ab5103-24.jpg)
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Product Name
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Histone H3 (citrulline 2 + 8 + 17) antibody [CitH3] - ChIP Grade
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See all Histone H3 (Citrullated) antibodies (4)...
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Product type
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Primary antibodies
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Description
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Rabbit polyclonal [CitH3] to Histone H3 (citrulline 2 + 8 + 17) - ChIP Grade
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Immunogen
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Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human Histone H3, at Cit2, 8 and 17. (Peptide available as ab32876.)
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Reacts with
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Hu, Ms, Cow, Mk, Rb
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Predicted to react
(species key)
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Rat, Many
(Due to sequence homology)
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Tested applications
(see key)
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ChIP, ChIP/Chip, ICC/IF, WB
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Abreviews
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Application notes
(see key)
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Recommended dilutions
ChIP: Use 6µl for 10 cells.
ChIP/Chip: Use at an assay dependent dilution.
IF: 1/250.
WB: 1/2000. Detects a band of approximately 15-17 kDa (predicted molecular weight: 15 kDa).
Not yet tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.
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Positive control
(see definition)
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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.
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Cellular localization
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Nuclear
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Research areas
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Chromatin and Nuclear Signaling >> ChIP'ing antibodies >> ChIP'ing antibodies
Chromatin and Nuclear Signaling >> Histones >> H3 >> Citrullated
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Relevance
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Histones 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).
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Alternative names Histone H3 (citrulline 2 + 8 + 17) antibody [CitH3] - ChIP Grade (ab5103) |
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Database links
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The links below go to external sites and will open in a new browser window |
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Raised in
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Rabbit
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Clonality
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Polyclonal
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Clone number
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CitH3
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Purity
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Immunogen affinity purified
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Storage buffer
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Preservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4 Material safety datasheet (MSDS) for this product:
Sodium Azide MSDS
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Form
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Liquid
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Concentration
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Batch dependent within range: 0.30 - 0.70 mg/ml
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Storage instructions
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Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
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At Abcam, we have one centralized database to hold all of our product information, so that everything we know about this Histone H3 (citrulline 2 + 8 + 17) antibody [CitH3] - ChIP Grade is on this datasheet. But please do contact us if you would like any reassurance! |
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See below for Histone H3 (citrulline 2 + 8 + 17) antibody [CitH3] - ChIP Grade images, references, products related to ab5103 and other tools.
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Histone H3 (citrulline 2 + 8 + 17) antibody [CitH3] - ChIP Grade images:
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 - Histone H3 (citrulline 2 + 8 + 17) antibody [CitH3] - ChIP Grade (ab5103)
Chromatin immunoprecipitation using ab5103 on the pS2 promoter. Times are after stimulation by estrogen (Ul). Cuthbert, G.L. et al, (2004) Cell 118, 545-553
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Immunocytochemistry/ Immunofluorescence - Histone H3 (citrulline 2 + 8 + 17) antibody [CitH3] - ChIP Grade (ab5103)
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).
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Western blot - Histone H3 (citrulline 2 + 8 + 17) antibody [CitH3] - ChIP Grade (ab5103)
All lanes : Histone H3 (citrulline 2 + 8 + 17) antibody [CitH3] - ChIP Grade (ab5103) at 1 µg/ml
Lane 1 : Calf Thymus Histone Preparation Nuclear Lysate at 0.5 µg
Lane 2 : HL60 (Human Caucasian promyelocytic leukaemia) Untreated Whole Cell Lysate at 10 µg
Lane 3 : HL60 (Human Caucasian promyelocytic leukaemia) DMSO and Calcium Lonophore treated Whole Cell Lysate at 10 µg
Lane 4 : Calf Thymus Histone Preparation Nuclear Lysate at 0.5 µg with Histone H3 peptide - citrulline 2 + 8 + 17 (ab32876) at 1 µg/ml
Lane 5 : HL60 (Human Caucasian promyelocytic leukaemia) Untreated Whole Cell Lysate at 10 µg with Histone H3 peptide - citrulline 2 + 8 + 17 (ab32876) at 1 µg/ml
Lane 6 : HL60 (Human Caucasian promyelocytic leukaemia) DMSO and Calcium Lonophore treated Whole Cell Lysate at 10 µg with Histone H3 peptide - citrulline 2 + 8 + 17 (ab32876) at 1 µg/ml
Lane 7 : Calf Thymus Histone Preparation Nuclear Lysate at 0.5 µg with Histone H3 peptide at 1 µg/ml
Lane 8 : HL60 (Human Caucasian promyelocytic leukaemia) Untreated Whole Cell Lysate at 10 µg with Histone H3 peptide at 1 µg/ml
Lane 9 : HL60 (Human Caucasian promyelocytic leukaemia) DMSO and Calcium Lonophore treated Whole Cell Lysate at 10 µg with Histone H3 peptide at 1 µg/ml
Secondary
Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 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?)
Exposure time : 30 seconds
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- Histone H3 (citrulline 17 + 2 + 8) antibody [CitH3] - ChIP Grade (ab5103)
![- Histone H3 (citrulline 17 + 2 + 8) antibody [CitH3] - ChIP Grade (ab5103)](/ps/datasheet/images/ab5103_2.jpg)
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.
Cuthbert, G.L. et al, (2004) Cell 118, 545-553
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References for Histone H3 (citrulline 2 + 8 + 17) antibody [CitH3] - ChIP Grade (ab5103)
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This product has been used in: (two most recent references)
Wang Y et al. Histone hypercitrullination mediates chromatin decondensation and neutrophil extracellular trap formation. J Cell Biol 184:205-13 (2009). WB; Human. PubMed: 19153223
Denis H et al. Functional connection between deimination and deacetylation of histones. Mol Cell Biol 29:4982-93 (2009). WB, ChIP/Chip. PubMed: 19581286
See all 8 publication references for this product.
If you publish research using ab5103 please let us know so that we can cite the reference on this datasheet.
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Search PubMed (MEDLINE) for references to Histone H3
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Histone H3 (citrulline 2 + 8 + 17) antibody [CitH3] - ChIP Grade - more information
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Histone H3 (citrulline 2 + 8 + 17) antibody [CitH3] - ChIP Grade - related products:
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Histone H3 (citrulline 2 + 8 + 17) antibody [CitH3] - ChIP Grade - other tools:
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All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
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![- Histone H3 (citrulline 2 + 8 + 17) antibody [CitH3] - ChIP Grade (ab5103)](/index.html?datasheet=5103#ab5103_1.jpg)
![Immunocytochemistry/ Immunofluorescence - Histone H3 (citrulline 2 + 8 + 17) antibody [CitH3] - ChIP Grade (ab5103)](/index.html?datasheet=5103#ab5103-IF-Im1.jpg)
![Western blot - Histone H3 (citrulline 2 + 8 + 17) antibody [CitH3] - ChIP Grade (ab5103)](/index.html?datasheet=5103#Histone-H3-Primary-antibodies-ab5103-24.jpg)
