Anti-Histone H1x antibody - ChIP Grade

4 product images

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  ChIP - Anti-Histone H1x antibody - ChIP Grade (ab31972)

  Chromatin was prepared from HeLa cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 minutes. The ChIP was performed with 25μg of chromatin, 2μg of ab31972 (blue), and 20μl of Protein A/G sepharose beads. No antibody was added to the beads control (yellow). The immunoprecipitated DNA was quantified by real time PCR (Taqman approach for active and inactive loci, Sybr green approach for heterochromatic loci). Primers and probes are located in the first kb of the transcribed region.

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  Western blot - Anti-Histone H1x antibody - ChIP Grade (ab31972)

  ab31972 recognises histone H1.X in HeLa nuclear extract at 35 kDa (lane 1), which is blocked using the immunizing peptide ab18052.

  All lanes: Western blot - Anti-Histone H1x antibody - ChIP Grade (ab31972) at 1 µg/mL

  Lane 1: HeLa nuclear lysate at 20 µg

  Lane 2: HeLa nuclear lysate at 20 µg with Human Histone H1x peptide (ab18052)

  Predicted band size: 22 kDa

  Observed band size: 35 kDa, 98 kDa

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H1x antibody - ChIP Grade (ab31972)

  IHC image of Histone H1x staining in Human breast adenocarcinoma formalin fixed paraffin embedded tissue section*, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab31972, 1μg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

  For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

  *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

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  Immunocytochemistry/ Immunofluorescence - Anti-Histone H1x antibody - ChIP Grade (ab31972)

  ICC/IF image of ab31972 stained human HeLa cells. The cells were PFA fixed (10 min), permabilised in TBS-T (20 min) and incubated with the antibody (ab31972, 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).