Anti-Histone H2B (mono methyl K5) antibody [EPR18092]

• IHC-P

AbReview78134****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H2B (mono methyl K5) antibody [EPR18092] (AB188340)

Immunohistochemistry analysis (Formalin/PFA-fixed paraffin-embedded sections) of formaldehyde fixed PBS-Tween permeabilized human colon tumor tissue. Stained with ab188340 at 1/2000 dilution. Secondary antibody used was HRP Agilent Detection System, Rabbit/Mouse. Blocking was done with 5% serum for 1 hour at 21°C. The sample was incubated with the primary antibody and TBST for 1 hour at 21°C. Antigen retrieval method was heat mediated Citrate

This image is courtesy of an anonymous Abreview

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Histone H2B (mono methyl K5) antibody [EPR18092] (AB188340)

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 K5) with ab188340 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 :
-ve control 1 : ab188340 at 1/2000 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
-ve 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

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H2B (mono methyl K5) antibody [EPR18092] (AB188340)

Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling Histone H2B (mono methyl K5) with ab188340 at 1/2000 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Nucleus staining on Human 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.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-Histone H2B (mono methyl K5) antibody [EPR18092] (AB188340)

Intracellular Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labelling Histone H2B (red) with purified ab188340 at a dilution of 1/250. The secondary antibody used was Alexa Fluorr^® 488 goat-anti-rabbit IgG (1/2000). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Isotype control antibody was rabbit monoclonal IgG (black). The blue line shows cells without incubation with primary and secondary antibody.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H2B (mono methyl K5) antibody [EPR18092] (AB188340)

Immunohistochemical analysis of paraffin-embedded Mouse colon tissue labeling Histone H2B (mono methyl K5) with ab188340 at 1/2000 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Nucleus 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.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H2B (mono methyl K5) antibody [EPR18092] (AB188340)

Immunohistochemical analysis of paraffin-embedded Rat colon tissue labeling Histone H2B (mono methyl K5) with ab188340 at 1/2000 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Nucleus staining on rat 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

Unknown

Western blot - Anti-Histone H2B (mono methyl K5) antibody [EPR18092] (AB188340)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Histone H2B (mono methyl K5) antibody [EPR18092] (ab188340) at 1/5000 dilution

Lane 1:

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

Lane 2:

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

Exposure time: 15s

• PepArr

Unknown

Peptide Array - Anti-Histone H2B (mono methyl K5) antibody [EPR18092] (AB188340)

ab188340 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.