Products:Epigenetics and Nuclear Signaling >> Histones >> H3 >> Citrullated
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ab46666 |
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ab5103 has been referenced in 9 publications.
Publishing research using ab5103? Please let us know so that we can cite the reference in this datasheet
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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
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).
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
Lane 5 : HL60 (Human Caucasian promyelocytic leukaemia) Untreated Whole Cell Lysate at 10 µg with
Lane 6 : HL60 (Human Caucasian promyelocytic leukaemia) DMSO and Calcium Lonophore treated Whole Cell Lysate at 10 µg with
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
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 25ºC. The primary antibody was diluted 1/250 in PBS and incubated with the sample for 12 hours at 4ºC. 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.
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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