Anti-Histone H2B (mono methyl K116) antibody [EPR17700]

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Histone H2B (mono methyl K116) antibody [EPR17700] (AB177787)

Immunofluorescent analysis of 4% paraformaldehyde-fixed 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling Histone H2B (mono methyl K116) with ab177787 at 1/2000 dilution followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green). Confocal image showing nuclear staining on HeLa cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).
The negative controls are as follows :
Negative control 1 : ab177787 at 1/2000 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
Negative control 2 : ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500 dilution.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H2B (mono methyl K116) antibody [EPR17700] (AB177787)

Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling Histone H2B (mono methyl K116) with ab177787 at 1/2000 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary at 1/500 dilution. Nucleus staining on glandular epithelium of human colon tissue is observed. Counter stained with Hematoxylin.

Negative control : Using 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

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H2B (mono methyl K116) antibody [EPR17700] (AB177787)

Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling Histone H2B (mono methyl K116) with ab177787 at 1/2000 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary at 1/500 dilution. Nucleus staining on mouse liver tissue is observed. Counter stained with Hematoxylin.

Negative control : Using 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

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H2B (mono methyl K116) antibody [EPR17700] (AB177787)

Immunohistochemical analysis of paraffin-embedded Rat pancreas tissue labeling Histone H2B (mono methyl K116) with ab177787 at 1/2000 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary at 1/500 dilution. Nucleus staining on rat pancreas tissue is observed. Counter stained with Hematoxylin.

Negative control : Using 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 H2B (mono methyl K116) antibody [EPR17700] (AB177787)

Blocking/Dilution buffer : 5% BSA/TBST.

ab177787 detects in HeLa cells a weaker signal than in NIH/3T3 cells. In the HeLa WB, a 5-fold more sensitive ECL substrate was used.

All lanes:

Western blot - Anti-Histone H2B (mono methyl K116) antibody [EPR17700] (ab177787) at 1/5000 dilution

All lanes:

HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate at 10 µg

Secondary

All lanes:

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

Predicted band size: 14 kDa

Observed band size: 14 kDa

false

• WB

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Western blot - Anti-Histone H2B (mono methyl K116) antibody [EPR17700] (AB177787)

Blocking/Dilution buffer : 5% BSA/TBST.

All lanes:

Western blot - Anti-Histone H2B (mono methyl K116) antibody [EPR17700] (ab177787) at 1/5000 dilution

All lanes:

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

Secondary

All lanes:

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

Predicted band size: 14 kDa

Observed band size: 14 kDa

false

• PepArr

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Peptide Array - Anti-Histone H2B (mono methyl K116) antibody [EPR17700] (AB177787)

ab177787 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 area under 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.