Anti-Histone H3 (citrulline R2 + R8 + R17) antibody - ChIP Grade

• Supplier: Abcam
• Catalog: ab5103
• Prices: $403.00
• Sizes: 100 µg
• Host: Rabbit
• Clonality: Polyclonal
• Isotype: IgG
• Applications: ICC/IF ICC/IF ICC/IF Array IHC-F ChIP ChIP WB ChIP IHC-P FC
• Species Reactivities: Mouse, Rat, Rabbit, Bovine, Human, C. elegans, Drosophila, Monkey, Broad species
• Antigen: Synthetic peptide corresponding to Human Histone H3 aa 1-100 (citrulline R2 + R8 + R17) conjugated to Keyhole Limpet Haemocyanin (KLH)
• Blocking Peptide: Human Histone H3 (citrulline R2 + R8 + R17) peptide
• Description: Rabbit Polyclonal
• Gene: CELE_F58A4.3
• Conjugate: Unconjugated

Product images

Anti-Histone H3 (citrulline R2 + R8 + R17) antibody - ChIP Grade (ab5103) at 1/1000 dilution + Fruit fly (Drosophila melanogaster) tissue lysate - nuclear at 5 µgSecondaryHRP-conjugated Goat anti-rabbit IgG polyclonal at 1/1000 dilutiondeveloped using the ECL techniquePerformed under reducing conditions.

Anti-Histone H3 (citrulline R2 + R8 + R17) antibody - ChIP Grade (ab5103) at 1/1000 dilution + Caenorhabditis elegans embryo at 30 µgSecondaryHRP-conjugated Goat anti-rabbit IgG polyclonal (undiluted)developed using the ECL techniquePerformed under reducing conditions.

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 + 17ab17566 - Histone H3 - unmodified

All lanes : Anti-Histone H3 (citrulline R2 + R8 + R17) antibody - ChIP Grade (ab5103) at 1 µg/mlLane 1 : HL60 (Human Caucasian promyelocytic leukaemia) DMSO and Calcium Ionophore treated Whole Cell Lysate with with 5% BSALane 2 : HL60 (Human Caucasian promyelocytic leukaemia) DMSO and Calcium Ionophore treated Whole Cell Lysate with with 5% milkLane 3 : HL60 (Human Caucasian promyelocytic leukaemia) DMSO and Calcium Ionophore treated Whole Cell Lysate with with 3% milkLysates/proteins at 10 µg per lane.SecondaryGoat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilutiondeveloped using the ECL techniquePerformed under reducing conditions.

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.See Abreview

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