Anti-Histone H1.0 antibody [EPR6537]

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Histone H1.0 antibody [EPR6537] (AB125027)

Immunofluorescence staining of BxPC-3 cells with purified ab125027 at a working dilution of 1/100, counter-stained with DAPI. The secondary antibody was Alexa Fluor^® 488 goat anti-rabbit (ab150077), used at a dilution of 1/1000. ab7291, a mouse anti-tubulin antibody (1/1000), was used to stain tubulin along with ab150120 (Alexa Fluor^® 594 goat anti-mouse, 1/1000), shown in the top right hand panel. The cells were fixed in 100% methanol and permeabilized using 0.1% Triton X 100. The negative controls are shown in bottom middle and right hand panels - for negative control 1, purified ab125027 was used at a dilution of 1/500 followed by an Alexa Fluor^® 594 goat anti-mouse antibody (ab150120) at a dilution of 1/500. For negative control 2, ab7291 (mouse anti-tubulin) was used at a dilution of 1/500 followed by an Alexa Fluor^® 488 goat anti-rabbit antibody (ab150077) at a dilution of 1/400.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H1.0 antibody [EPR6537] (AB125027)

Immunohistochemical staining of paraffin embedded human thyroid carcinoma with purified ab125027 at a working dilution of 1/400. The secondary antibody used is HRP goat anti-rabbit IgG H&L (ab97051) at 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Histone H1.0 antibody [EPR6537] (AB125027)

Unpurified ab125027, at a 1/100 dilution, staining Histone H1 in A431 cells by Immunofluorescence.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H1.0 antibody [EPR6537] (AB125027)

Immunohistochemical staining of paraffin embedded human pancreas with purified ab125027 at a working dilution of 1/400. The secondary antibody used is HRP goat anti-rabbit IgG H&L (ab97051) at 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H1.0 antibody [EPR6537] (AB125027)

Unpurified ab125027, at a 1/100 dilution, staining Histone H1 in paraffin embedded Human kidney tissue by Immunohistochemistry.

• WB

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Western blot - Anti-Histone H1.0 antibody [EPR6537] (AB125027)

All lanes:

Western blot - Anti-Histone H1.0 antibody [EPR6537] (ab125027) at 1/1000 dilution

Lane 1:

BxPC3 lysates at 10 µg

Lane 2:

Human fetal lung lysates at 10 µg

Secondary

All lanes:

HRP labelled goat anti rabbit at 1/2000 dilution

Predicted band size: 21 kDa

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• WB

Lab

Western blot - Anti-Histone H1.0 antibody [EPR6537] (AB125027)

Lanes 1 - 2 : Merged signal (red and green). Green - ab125027 observed at 33 kDa. Red - loading control, ab7291 (Mouse anti-Alpha Tubulin [DM1A] observed at 55kDa.

ab125027 was shown to react with H1F0 in A431 wild-type cells in Western blot. Loss of signal was observed when H1F0 knockout sample was used. A431 wild-type 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 ab125027 and ab7291 (Mouse anti-Alpha Tubulin [DM1A] overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Histone H1.0 antibody [EPR6537] (ab125027) at 1/1000 dilution

Lane 1:

Wild-type A431 (Human epidermoid carcinoma cell line) whole cell lysate at 20 µg

Lane 2:

H1F0 knockout A431 (Human epidermoid carcinoma cell line) whole cell lysate at 20 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>)

Predicted band size: 21 kDa

Observed band size: 33 kDa

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• WB

Lab

Western blot - Anti-Histone H1.0 antibody [EPR6537] (AB125027)

Blocking buffer : 5% NFDM/TBST
Dilution buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-Histone H1.0 antibody [EPR6537] (ab125027) at 10 µg

Lane 1:

human fetal lung lysate

Lane 2:

BxPC-3 lysate

Lane 3:

A431 lysate

Lane 4:

human fetal kidney lysate

Secondary

All lanes:

HRP goat anti-rabbit IgG (H+L) at 1/1000 dilution

Predicted band size: 106 kDa,110 kDa,118 kDa,150 kDa,21 kDa,30 kDa,50 kDa,64 kDa,84 kDa

Observed band size: 30 kDa,32 kDa,65 kDa,80 kDa

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