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

Overview

• Product nameAnti-Histone H3 (citrulline 2 + 8 + 17) antibody - ChIP GradeSee all Histone H3 primary antibodies ...
• Description
  Rabbit polyclonal to Histone H3 (citrulline 2 + 8 + 17) - ChIP Grade
• Tested applicationsICC/IF, IHC-Fr, ChIP/Chip, WB, ChIP, IHC-P more details
• Species 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 controlThis 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

• FormLiquid
• 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
  • Epigenetics and Nuclear Signaling
  • ChIP'ing antibodies
  • ChIP'ing antibodies

  • Epigenetics and Nuclear Signaling
  • Histones
  • H3
  • Citrullated

Applications

Target

• 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
• Database links
  • Entrez Gene: 326601 Cow
  • Entrez Gene: 3020 Human
  • Entrez Gene: 3021 Human
  • Entrez Gene: 10338 Human
  • Entrez Gene: 15079 Mouse
  • Entrez Gene: 15080 Mouse
  • Entrez Gene: 289314 Rat
  • Entrez Gene: 117056 Rat

  • Omim: 601128 Human
  • SwissProt: P16106 Cow
  • SwissProt: P16105 Cow
  • SwissProt: P16106 Human
  • SwissProt: P06351 Human
  • SwissProt: Q93081 Human
  • SwissProt: Q16695 Human
  • SwissProt: P16105 Mouse
  • SwissProt: P84244 Mouse
  • SwissProt: A2A852 Mouse
  • SwissProt: P06351 Rat
  • SwissProt: P84245 Rat
  • SwissProt: B0BMY8 Rat
  • Unigene: 181307 Human

  see all

• Alternative names
  Cit Histone H3 antibodyCitrullated antibodycitrullin antibody

  Citrullinated Histone H3 antibodycitrulline antibodyH3 3 like sequence MH921 antibodyH3 3A antibodyH3 histone antibodyH3 histone family member E pseudogene antibodyH3F3 antibodyHIST3H3 antibodyHistone H3 3 pseudogene antibodyHistone H3 Citrullated antibody

  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)Cuthbert, G.L. et al, (2004) Cell 118, 545-553
  Chromatin immunoprecipitation using ab5103 on the pS2 promoter. Times are after stimulation by estrogen (Ul).
• 

  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)
  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)This image is courtesy of an Anonymous Abreview.
  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.

  See Abreview

• 

  Western blot - Anti-Histone H3 (citrulline 2 + 8 + 17) antibody - ChIP Grade (ab5103)
  Lanes 1 - 3 : Anti-Histone H3 (citrulline 2 + 8 + 17) antibody - ChIP Grade (ab5103) at 1 µg/ml
  Lanes 4 - 6 : Anti-Histone H3 (citrulline 2 + 8 + 17) antibody - ChIP Grade (ab5103) at 1 µg/ml (5% Milk Block)
  Lanes 7 - 9 : Anti-Histone H3 (citrulline 2 + 8 + 17) antibody - ChIP Grade (ab5103) at 1 µg/ml (3% Milk Block)
  
  Lane 1 : HL60 (Human Caucasian promyelocytic leukaemia) DMSO and Calcium Lonophore treated Whole Cell Lysate
  Lane 2 : HL60 (Human Caucasian promyelocytic leukaemia) DMSO and Calcium Lonophore treated Whole Cell Lysate with Histone H3 peptide - citrulline 2 + 8 + 17 (ab32876) at 1 µg/ml
  Lane 3 : HL60 (Human Caucasian promyelocytic leukaemia) DMSO and Calcium Lonophore treated Whole Cell Lysate with Histone H3 peptide at 1 µg/ml
  Lane 4 : HL60 (Human Caucasian promyelocytic leukaemia) DMSO and Calcium Lonophore treated Whole Cell Lysate
  Lane 5 : HL60 (Human Caucasian promyelocytic leukaemia) DMSO and Calcium Lonophore treated Whole Cell Lysate 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 with Histone H3 peptide at 1 µg/ml
  Lane 7 : HL60 (Human Caucasian promyelocytic leukaemia) DMSO and Calcium Lonophore treated Whole Cell Lysate
  Lane 8 : HL60 (Human Caucasian promyelocytic leukaemia) DMSO and Calcium Lonophore treated Whole Cell Lysate with Histone H3 peptide - citrulline 2 + 8 + 17 (ab32876) at 1 µg/ml
  Lane 9 : HL60 (Human Caucasian promyelocytic leukaemia) DMSO and Calcium Lonophore treated Whole Cell Lysate with Histone H3 peptide at 1 µg/ml
  
  Lysates/proteins at 10 µg per lane.
  
  Secondary
  IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) (ab28446) 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
• 

  - Histone H3 (citrulline 2 + 8 + 17) antibody [CitH3] - ChIP Grade (ab5103)Cuthbert, G.L. et al, (2004) Cell 118, 545-553

  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.