Anti-Histone H1.0 antibody [34]

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H1.0 antibody [34] (AB11079)

IHC image of Histone H1.0 staining in a formalin fixed, paraffin embedded human breast adenocarcinama 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 ab11079, 5μg/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

• WB

Collaborator

Western blot - Anti-Histone H1.0 antibody [34] (AB11079)

Mouse monoclonal to Histone H1.0 against recombinant human H1 isoforms H1.0 to H1.5 as depicted in the figure.

Dilution 1/1000.

Recombinant H1 isoforms courtesy of Dr Nicole Happel.

(see attached review)

All lanes:

Western blot - Anti-Histone H1.0 antibody [34] (ab11079)

Predicted band size: 21 kDa

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Dr Albert Jordan, Center for Genomic Regulation (CRG), Spain.

• WB

Unknown

Western blot - Anti-Histone H1.0 antibody [34] (AB11079)

Western blot using ab11079 on HeLa whole cell extract (30 μg/lane).

Exposure 5 sec using ECL.
12.5% SDS-PAGE gel.

All lanes:

Western blot - Anti-Histone H1.0 antibody [34] (ab11079)

Predicted band size: 21 kDa

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• WB

Lab

Western blot - Anti-Histone H1.0 antibody [34] (AB11079)

Lanes 1 - 3 : Merged signal (red and green). Green - ab11079 observed at 30 kDa. Red - loading control ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) observed at 55 kDa.

ab11079 was shown to react with Histone H1.0 in wild-type A431 cells in Western blot with loss of signal observed in H1F0 knockout sample. Wild-type A431 and H1F0 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween^®) before incubation with ab11079 and ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) overnight at 4°C at a 1 in 500 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Mouse IgG H&L (IRDye^® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye^® 680RD) preabsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.

All lanes:

Western blot - Anti-Histone H1.0 antibody [34] (ab11079) at 1/500 dilution

Lane 1:

Wild-type A431 cell lysate at 40 µg

Lane 2:

H1F0 knockout A431 cell lysate at 40 µg

Lane 2:

Western blot - Human H1F0 knockout A-431 cell line (<a href='/en-us/products/cell-lines/human-h1f0-knockout-a-431-cell-line-ab262478'>ab262478</a>)

Lane 3:

MCF7 (Human breast adenocarcinoma cell line) whole cell lysate at 40 µg

Predicted band size: 21 kDa

Observed band size: 30 kDa

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• WB

Lab

Western blot - Anti-Histone H1.0 antibody [34] (AB11079)

All lanes:

Western blot - Anti-Histone H1.0 antibody [34] (ab11079) at 1 µg/mL

Lane 1:

Histone H1.0 (Human) - Recombinant Protein at 0.1 µg

Lane 2:

Histone H1.2 (Human) - Recombinant Protein at 0.1 µg

Secondary

All lanes:

Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) (ab65485) at 1/5000 dilution

Predicted band size: 21 kDa

Observed band size: 32 kDa

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Exposure time: 10s

• WB

Lab

Western blot - Anti-Histone H1.0 antibody [34] (AB11079)

All lanes:

Western blot - Anti-Histone H1.0 antibody [34] (ab11079) at 1 µg/mL

All lanes:

HeLa (Triton-enriched) Cell Lysate at 10 µg

Secondary

All lanes:

Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) (ab65485) at 1/5000 dilution

Predicted band size: 21 kDa

Observed band size: 120 kDa,18 kDa,32 kDa

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Exposure time: 8min

• WB

CiteAb

Western blot - Anti-Histone H1.0 antibody [34] (AB11079)

Histone H1.0 western blot using anti-Histone H1.0 antibody [34] ab11079. Publication image and figure legend from Paupe, V., Dassa, E. P., et al., 2009, PLoS One, PubMed 19158945.

ab11079 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab11079 please see the product overview.

Nrf2 location and amount in control and FRDA patient fibroblasts under basal and oxidative stress conditions.A. Nrf2 localization in control (a, b, c) and patient (d, e, f) fibroblasts under basal conditions (a, d) or after treatment with oligomycin (b, e) or tBHQ (c, f). Nuclear Nrf2 translocation occurred in control cells, but not patient cells, after oligomycin or tBHQ treatment. B. Western blots of cytoplasmic (a; 40 μg/lane protein) and nuclear (b; 60 μg/lane protein) fractions from control (lanes 1–3) and patient (lanes 4–6) fibroblasts under basal conditions (lane 1, 4), after oligomycin treatment (lane 2, 5), or after tBHQ treatment (lane 3, 6). The specificity of the 100 kDa band was confirmed using two antibodies (H-300 and C-20). Fraction purity (d) was assessed by labeling with GAPDH (cytoplasm) and Histone H1 antibody (nuclei). C. Nrf2 content relative to GAPDH and Histone H1 contents in cytoplasmic and nuclear fractions of control and patient fibroblasts. Asterisks denote significant differences (*p<0.05 and **p<0.01). Values are means±1 SEM. Experimental procedures are described in the methods section.

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