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Anti-Histone H3 (citrulline 2 + 8 + 17) antibody - ChIP Grade (ab5103)

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Overview

Product name

Anti-Histone H3 (citrulline 2 + 8 + 17) antibody - ChIP Grade
See all Histone H3 products (4) ...

Description

Rabbit polyclonal to Histone H3 (citrulline 2 + 8 + 17) - ChIP Grade

Tested applications

ICC/IF, IHC-Fr, ChIP/Chip, WB, ChIP, IFmore details

Cross reactivity

Reacts with

Mouse, Rat, Rabbit, Cow, Human, Monkey

Predicted to work with

a wide range of other species

Immunogen

Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human Histone H3, at Cit2, 8 and 17.

Read Abcam's proprietary immunogen policy

(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.

Properties

Form

Liquid

Storage instructions

Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.

Storage buffer

Preservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4

Concentration

Concentration information loading...

Purity

Immunogen affinity purified

Clonality

Polyclonal

Research areas

Epigenetics and Nuclear Signaling >> ChIP'ing antibodies >> ChIP'ing antibodies
Epigenetics and Nuclear Signaling >> Histones >> H3 >> Citrullated

  • ChIP - Histone H3 (citrulline 2 + 8 + 17) antibody [CitH3] - ChIP Grade (ab5103)ChIP - Histone H3 (citrulline 2 + 8 + 17) antibody [CitH3] - ChIP Grade (ab5103) image (enlarge)

  • 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) image (enlarge)

  • 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) image (enlarge)

Applications

Show applications key

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.

Target

Relevance

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).

Cellular localization

Nuclear

Alternative names

  • 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
see all

Database links

see all

Anti-Histone H3 (citrulline 2 + 8 + 17) antibody - ChIP Grade images:

  ChIP - Histone H3 (citrulline 2 + 8 + 17) antibody [CitH3] - ChIP Grade (ab5103)

ChIP - 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

  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)

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).

  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)

All lanes : Anti-Histone H3 (citrulline 2 + 8 + 17) antibody - 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

  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)

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.

This image is courtesy of an Anonymous Abreview.

See Abreview

  - Histone H3 (citrulline 2 + 8 + 17) antibody [CitH3] - ChIP Grade (ab5103)

- Histone H3 (citrulline 2 + 8 + 17) antibody [CitH3] - ChIP Grade (ab5103)

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

References for Anti-Histone H3 (citrulline 2 + 8 + 17) antibody - ChIP Grade (ab5103)

This product has been referenced in:

  • Douda DNet al. Innate immune collectin surfactant protein d simultaneously binds both neutrophil extracellular traps and carbohydrate ligands and promotes bacterial trapping. J Immunol 187:1856-65 (2011). WB; Mouse.Read more (PubMed: 21724991) »
  • Li Pet al. PAD4 is essential for antibacterial innate immunity mediated by neutrophil extracellular traps. J Exp Med 207:1853-62 (2010). WB, ICC/IF; Mouse.Read more (PubMed: 20733033) »

See all 9 publications for this product

Publishing research using ab5103? Please let us know so that we can cite the reference in this datasheet

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"