Anti-Histone H2B (mono methyl K85) antibody [EPR17613] - BSA and Azide free

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Histone H2B (mono methyl K85) antibody [EPR17613] - BSA and Azide free (AB249993)

This data was developed using ab177781, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (human epithelial cell line from cervix adenocarcinoma) cells labeling Histone H2B (mono methyl K85) with ab177781 at 1/2000 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear staining on HeLa cell line.

The nuclear counter stain is DAPI (blue).

Tubulin is detected with Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) (ab150120) secondary antibody at 1/500 dilution (red).

The negative controls are as follows :

-ve control 1 : ab177781 at 1/2000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) (ab150120) secondary antibody at 1/500 dilution.

-ve control 2 : Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/500 dilution.

• WB

Supplier Data

Western blot - Anti-Histone H2B (mono methyl K85) antibody [EPR17613] - BSA and Azide free (AB249993)

This data was developed using ab177781, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% BSA/TBST.

All lanes:

Western blot - Anti-Histone H2B (mono methyl K85) antibody [EPR17613] (<a href='/en-us/products/primary-antibodies/histone-h2b-mono-methyl-k85-antibody-epr17613-ab177781'>ab177781</a>) at 1/5000 dilution

All lanes:

HL-60 (human promyelocytic leukemia cell line) cell lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/50000 dilution

Predicted band size: 14 kDa

Observed band size: 14 kDa

true

Exposure time: 5s

• WB

Supplier Data

Western blot - Anti-Histone H2B (mono methyl K85) antibody [EPR17613] - BSA and Azide free (AB249993)

This data was developed using ab177781, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% BSA/TBST.

Under our testing conditions (using whole cell extract and BSA blocking) we detected non-specific bands in addition to the BOI.

All lanes:

Western blot - Anti-Histone H2B (mono methyl K85) antibody [EPR17613] (<a href='/en-us/products/primary-antibodies/histone-h2b-mono-methyl-k85-antibody-epr17613-ab177781'>ab177781</a>) at 1/1000 dilution

Lane 1:

Human ovary lysate at 20 µg

Lane 2:

HeLa (human epithelial cell line from cervix adenocarcinoma) cell lysate at 20 µg

Lane 3:

NIH:OVCAR-3 (human ovary adenocarcinoma cell line) cell lysate at 20 µg

Lane 4:

MOLT-4 (human lymphoblastic leukemia cell line) cell lysate at 20 µg

Lane 5:

THP-1 (human monocytic leukemia cell line) cell lysate at 20 µg

Lane 6:

K562 (human chronic myelogenous leukemia cell line from bone marrow) cell lysate at 20 µg

Lane 7:

F9 (mouse embryonic testicular cancer cell line) cell lysate at 20 µg

Lane 8:

RAW 264.7 (mouse macrophage cell line transformed with Abelson murine leukemia virus) cell lysate at 20 µg

Lane 9:

NIH/3T3 (mouse embyro fibroblast cell line) cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/50000 dilution

Predicted band size: 14 kDa

Observed band size: 14 kDa

true

Exposure time: 15s

• WB

Supplier Data

Western blot - Anti-Histone H2B (mono methyl K85) antibody [EPR17613] - BSA and Azide free (AB249993)

This data was developed using ab177781, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% BSA/TBST.

All lanes:

Western blot - Anti-Histone H2B (mono methyl K85) antibody [EPR17613] (<a href='/en-us/products/primary-antibodies/histone-h2b-mono-methyl-k85-antibody-epr17613-ab177781'>ab177781</a>) at 1/500 dilution

All lanes:

Mouse ovary tissue lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/50000 dilution

Predicted band size: 14 kDa

Observed band size: 14 kDa

true

Exposure time: 5s

• PepArr

Supplier Data

Peptide Array - Anti-Histone H2B (mono methyl K85) antibody [EPR17613] - BSA and Azide free (AB249993)

This data was developed using ab177781, the same antibody clone in a different buffer formulation.

ab177781 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.