Anti-Histone H3 (phospho S10) 抗体

• WB

Lab

Western blot - Anti-Histone H3 (phospho S10) antibody (AB5176)

All lanes:

Western blot - Anti-Histone H3 (phospho S10) antibody (ab5176) at 1/5000 dilution

Lane 1:

Untreated HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 10 µg

Lane 2:

HeLa cell lysate treated with calyculin A at 10 µg

Lane 3:

HeLa cell lysate treated with calyculin A and alkaline phosphatase at 10 µg

Secondary

All lanes:

HRP goat anti-rabbit (H+L) at 1/20000 dilution

Predicted band size: 15 kDa

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Exposure time: 1s

• ICC

Collaborator

Immunocytochemistry - Anti-Histone H3 (phospho S10) antibody (AB5176)

This image was kindly supplied by Prof Bryan Turner, University of Birmingham. Female Human Lymphoblastoid cells were incubated with ab5176.

Image courtesy of Dr. Bryan Turner, United Kingdom

• ICC

AbReview4926****

Immunocytochemistry - Anti-Histone H3 (phospho S10) antibody (AB5176)

ab5176 at a 1/100 dilution staining Drosophila melanogaster (wild type) polytene chromosomes by ICC/IF. The cells were formaldehyde fixed and blocked with 1% BSA prior to incubation with the antibody for 12 hours. Bound antibody was detected using a Cy3 conjugated goat anti-rabbit antibody.

This image is courtesy of an Abreview submitted by Miss Anita Ciurciu

• ICC

PubMed

Immunocytochemistry - Anti-Histone H3 (phospho S10) antibody (AB5176)

Images of mouse cardiomyocyte in various stages of mitosis. Puromycin-resistant cells were stained with Hoechst 33342 (blue), and probed with antibodies specific to cTnT (Green) and Histone H3 (phospho S10) (Red) using ab5176 at 1/200 dilution in ICC/IF.

Mitotic activity was confirmed by staining with an antibody to phospho-H3 Histone, which is only phosphorylated at Ser10 during mitosis. Mitotic nuclei are pink (red+blue). Bar =10 μm.

Yamanaka S et al. Enhanced proliferation of monolayer cultures of embryonic stem (ES) cell-derived cardiomyocytes following acute loss of retinoblastoma. PLoS ONE 3:e3896 (2008).

• WB

Unknown

Western blot - Anti-Histone H3 (phospho S10) antibody (AB5176)

All lanes:

Western blot - Anti-Histone H3 (phospho S10) antibody (ab5176) at 1 µg/mL

Lane 1:

untreated histones at 0.5 µg

Lane 2:

colcemid treated histones at 0.5 µg

Lane 3:

untreated histones at 0.5 µg with Human Histone H3 (phospho S10) peptide (<a href='/products/proteins-peptides/human-histone-h3-phospho-s10-peptide-ab11477'>ab11477</a>)

Lane 4:

colcemid treated histones at 0.5 µg with Human Histone H3 (phospho S10) peptide (<a href='/products/proteins-peptides/human-histone-h3-phospho-s10-peptide-ab11477'>ab11477</a>)

Lane 5:

untreated histones at 0.5 µg with Human Histone H3 (unmodified ) peptide (<a href='/products/proteins-peptides/human-histone-h3-unmodified-peptide-ab2903'>ab2903</a>)

Lane 6:

colcemid treated histones at 0.5 µg with Human Histone H3 (unmodified ) peptide (<a href='/products/proteins-peptides/human-histone-h3-unmodified-peptide-ab2903'>ab2903</a>)

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab6721'>ab6721</a>) at 1/5000 dilution

Predicted band size: 15 kDa

Observed band size: 15-20 kDa

true

Exposure time: 10s

• WB

CiteAb

Western blot - Anti-Histone H3 (phospho S10) antibody (AB5176)

Western Blotting using Anti-Histone H3 (phospho S10) antibody, ab5176. Publication image from Li, Z. et al., 2018, Cell Res, 29844578. Legend direct from paper.

H1.2 PARylation permits its displacement from chromatin upon DNA damage. a HeLa cells were transfected with GFP-H1.2 and treated with 20 µM Ku55933 or 2 µM Ku57788 for 4 h or 5 µM PJ34 for 1 h followed by laser micro-irradiation. Images were taken every 10 s for 5 min and quantifications of the IR path signal intensity were shown and ~15 IR paths from 10 separate cells were calculated. The data represent the mean ± SD. Scale bars, 10 µm. b HeLa cells were transfected with the indicated siRNAs and treated with 40 µM etoposide for the indicated time. Chromatin was fractionated and analyzed by immunoblotting. cParp1 wild-type (+/+) or KO (−/−) MEFs were treated with 40 µM etoposide for the indicated time and chromatin was fractionated and analyzed by immunoblotting. d HeLa cells were transfected with FLAG-H1.2 and treated with 40 µM etoposide for 15 min with or without 5 µM PJ34 for 1 h. Cell extracts were immunoprecipitated with FLAG-conjugated M2 beads. e Recombinant HIS-H1.2 was subjected to in vitro PARylation assay in the presence of NAD+ or 10 µM PJ34, as indicated. f HeLa cells were transfected with wild-type or S188A mutated FLAG-H1.2 and treated with 40 µM etoposide for 15 min with or without 5 µM PJ34 for 1 h, as indicated. Cells were extracted and immunoprecipitated with FLAG-conjugated M2 beads. g Recombinant wild-type, S188A mutated or C1-deleted (δC1) HIS-H1.2 were subjected to in vitro PARylation assay. h HeLa cells were transfected with wild-type, δC1 or S188A mutated GFP-H1.2 and subjected to laser micro-irradiation. Images were taken every 20 s for 5 min and representative images were shown. Quantifications were calculated as in a. The data represent the mean ± SD. Scale bars, 10 µm

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