Anti-Histone H3 (mono methyl R2) antibody [EPR17704] - BSA and Azide free

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (mono methyl R2) antibody [EPR17704] - BSA and Azide free (AB249932)

This data was developed using ab176844, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling Histone H3 (mono methyl R2) with ab176844 at 1/1000 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 (Alexa Fluor^® 594 Goat anti-Mouse secondary) at 1/500 dilution (red).
The negative controls are as follows :
-ve control 1 : ab176844 at 1/1000 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.

• ChIP

Supplier Data

ChIP - Anti-Histone H3 (mono methyl R2) antibody [EPR17704] - BSA and Azide free (AB249932)

This data was developed using ab176844, the same antibody clone in a different buffer formulation.

Chromatin was prepared from HeLa (Human epithelial cells from cervix adenocarcinoma) cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 minutes. The ChIP was performed with 25μg of chromatin, 2μg of ab176844 (blue), and 20μl of Anti rabbit IgG sepharose beads. 2μg of rabbit normal IgG was added to the beads control (yellow). The immunoprecipitated DNA was quantified by real time PCR (Sybr green approach). Primers and probes are located in the first kb of the transcribed region.

• WB

Supplier Data

Western blot - Anti-Histone H3 (mono methyl R2) antibody [EPR17704] - BSA and Azide free (AB249932)

This data was developed using ab176844, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% BSA /TBST.

All lanes:

Western blot - Anti-Histone H3 (mono methyl R2) antibody [EPR17704] - ChIP Grade (<a href='/en-us/products/primary-antibodies/histone-h3-mono-methyl-r2-antibody-epr17704-chip-grade-ab176844'>ab176844</a>) 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,15 kDa,30 kDa

Observed band size: 14 kDa,15 kDa,31 kDa

false

Exposure time: 15s

• WB

Supplier Data

Western blot - Anti-Histone H3 (mono methyl R2) antibody [EPR17704] - BSA and Azide free (AB249932)

This data was developed using ab176844, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% BSA /TBST.

All lanes:

Western blot - Anti-Histone H3 (mono methyl R2) antibody [EPR17704] - ChIP Grade (<a href='/en-us/products/primary-antibodies/histone-h3-mono-methyl-r2-antibody-epr17704-chip-grade-ab176844'>ab176844</a>) at 1/5000 dilution

All lanes:

NIH/3T3 (mouse embryo) whole 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: 30s

• PepArr

Supplier Data

Peptide Array - Anti-Histone H3 (mono methyl R2) antibody [EPR17704] - BSA and Azide free (AB249932)

This data was developed using ab176844, the same antibody clone in a different buffer formulation.ab176884 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.