Anti-Histone H3 antibody [EPR17785] - Nuclear Marker

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 antibody [EPR17785] - Nuclear Marker (AB201456)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton-X100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling Histone H3 with ab201456 at 1/800 dilution, followed by AlexaFluor®488 Goat anti-Rabbit secondary antibody (ab150077) at 1/500 dilution (green).

Confocal image showing nuclear staining on HeLa cell line.

The nuclear counter stain is DAPI (blue).

Tubulin is stained with ab7291 anti-Tubulin (mouse mAb) at 1/1000 dilution, followed by AlexaFluor®594 Goat anti-Mouse secondary antibody (ab150120) at 1/500 dilution (red).
-ve control 1 : ab201456 at 1/800 dilution followed by AlexaFluor®594 Goat anti-Mouse secondary antibody (ab150120) at 1/500 dilution.
-ve control 2 : ab7291 anti-Tubulin (mouse mAb) at 1/1000 dilution, followed by AlexaFluor®488 Goat anti-Rabbit secondary antibody (ab150077) at 1/500 dilution.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 antibody [EPR17785] - Nuclear Marker (AB201456)

Immunohistochemical analysis of paraffin-embedded Human cervix carcinoma tissue labeling Histone H3 with ab201456 at 1/200 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution.

Nuclear staining on Human cervix carcinoma tissue is observed.

Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 antibody [EPR17785] - Nuclear Marker (AB201456)

Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling Histone H3 with ab201456 at 1/200 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution.

Nuclear staining on Human kidney tissue is observed.

Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-Histone H3 antibody [EPR17785] - Nuclear Marker (AB201456)

Intracellular flow cytometric analysis of 2% paraformaldehyde-fixed HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling Histone H3with ab201456 at 1/100 dilution (red) compared with a rabbit monoclonal IgG isotype control (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (FITC) at 1/150 dilution was used as the secondary antibody.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 antibody [EPR17785] - Nuclear Marker (AB201456)

Immunohistochemical analysis of paraffin-embedded Rat stomach tissue labeling Histone H3 with ab201456 at 1/200 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution.

Nuclear staining on rat stomach tissue is observed.

Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 antibody [EPR17785] - Nuclear Marker (AB201456)

Immunohistochemical analysis of paraffin-embedded Mouse colon tissue labeling Histone H3 with ab201456 at 1/200 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution.

Nuclear staining on mouse colon tissue is observed.

Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• WB

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Western blot - Anti-Histone H3 antibody [EPR17785] - Nuclear Marker (AB201456)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Histone H3 antibody [EPR17785] - Nuclear Marker (ab201456) at 1/2000 dilution

Lane 1:

HeLa (Human epithelial cells from cervix adenocarcinoma) cell lysate at 20 µg

Lane 2:

HEK293 (Human embryonic kidney) cell lysate at 20 µg

Lane 3:

A375 (Human malignant melanoma) cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 15 kDa

Observed band size: 15 kDa

false

Exposure time: 15s

• WB

Lab

Western blot - Anti-Histone H3 antibody [EPR17785] - Nuclear Marker (AB201456)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

ab181602 was used as GAPDH loading control at a 1/2200000 dilution. ab201456 was used for total protein control.

All lanes:

Western blot - Anti-Histone H3 (phospho S28) antibody [HTA28] (<a href='/en-us/products/primary-antibodies/histone-h3-phospho-s28-antibody-hta28-ab10543'>ab10543</a>) at 1/1000 dilution

Lane 1:

Untreated HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate (untreated membrane) at 20 µg

Lane 2:

HeLa treated with 100ng/ml nocodazole for 16 hours whole cell lysate (untreated membrane) at 20 µg

Lane 3:

Untreated HeLa whole cell lysate (Lambda Protein Phosphatase treated membrane) at 20 µg

Lane 4:

HeLa treated with 100ng/ml nocodazole for 16 hours whole cell lysate (Lambda Protein Phosphatase treated membrane) at 20 µg

Secondary

All lanes:

Western blot at 1/10000 dilution

Predicted band size: 15 kDa

false

Exposure time: 15s

• WB

Supplier Data

Western blot - Anti-Histone H3 antibody [EPR17785] - Nuclear Marker (AB201456)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Histone H3 antibody [EPR17785] - Nuclear Marker (ab201456) at 1/2000 dilution

All lanes:

Human fetal brain lysate at 10 µg

Secondary

All lanes:

Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

Predicted band size: 15 kDa

Observed band size: 15 kDa

false

Exposure time: 3min

• WB

Supplier Data

Western blot - Anti-Histone H3 antibody [EPR17785] - Nuclear Marker (AB201456)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Histone H3 antibody [EPR17785] - Nuclear Marker (ab201456) at 1/2000 dilution

Lane 1:

Mouse kidney lysate at 10 µg

Lane 2:

Rat brain lysate at 10 µg

Lane 3:

NIH/3T3 (Mouse embyro fibroblast cells) cell lysate at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 15 kDa

Observed band size: 15 kDa

false

Exposure time: 1min

• PepArr

Lab

Peptide Array - Anti-Histone H3 antibody [EPR17785] - Nuclear Marker (AB201456)

Peptide array analysis of ab201456 at 1/10000 (0.02 ug/ml) dilution followed by a Goat Anti-Rabbit IgG, (H+L), Fluo 647nm conjugated at 1/50000 dilution.

Blocking buffer : 5% BSA in TBST.

ab201456 was tested in Peptide Array against 501 different modified and unmodified histone peptides; each peptide is printed on the array at six concentrations (each in triplicate).
Circle area represents affinity between the antibody and a peptide : all antigen-containing peptides are displayed as red circles, all other peptides as blue circles. The affinity is calculated as the area under the curve when antibody binding values are plotted against the corresponding peptide concentration. Each circle area is normalized to the peptide with the strongest affinity.

The complete dataset, including full list of all peptides and information on the position of each peptide in the diagram, is available under the Product Protocol section.

• WB

CiteAb

Western blot - Anti-Histone H3 antibody [EPR17785] - Nuclear Marker (AB201456)

Histone H3 western blot using anti-Histone H3 antibody [EPR17785] ab201456. Publication image and figure legend from Bitar, M. S., Nader, J., et al., 2018, Oxid Med Cell Longev, PubMed 30510624.

ab201456 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 ab201456 please see the product overview.

Diabetes alters key signaling molecules involved in skeletal muscle mass regulation. (a) Representative immunoblots and quantifications of phosphorylated Akt, S6, and 4E-BP1 protein levels in muscles of control and GK diabetic rats. (b) Total phosphorylated levels of FoxO1 and its nuclear localization relative to GAPDH and histone, respectively, were analyzed by Western blot. (c) Expression of MuRF1 and Atrogin 1 mRNAs relative to GAPDH was examined by real-time PCR. (d) Expression of miRNA 486 relative to U6 was determined using real-time PCR (e) PTEN protein level (e) and activity (f) were measured as described in Materials and Methods. (f) Key regulators of myogenesis including IGF-1 and PAX7 (g) were assessed in muscle using real-time RT-PCR-based technique. Abbreviation : C : control; D : diabetic. Values are means ± SEM for at least 6 animals/group. ∗Significantly different from corresponding control values at P ≤ 0.05.

false