Anti-Histone H3 (symmetric di methyl R26) antibody [EPR17719] - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (symmetric di methyl R26) antibody [EPR17719] - BSA and Azide free (AB249954)

This data was developed using ab177284, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded human stomach tissue labeling Histone H3 (symmetric di methyl R26) with ab177284 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear staining on human 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.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (symmetric di methyl R26) antibody [EPR17719] - BSA and Azide free (AB249954)

This data was developed using ab177284, 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 (symmetric di methyl R26) with ab177284 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear staining on HeLa cell line. The nuclear counterstain 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 at 1/1000 dilution (red).
The negative controls are as follows : -
-ve control 1 : ab177284 at 1/1000 dilution followed by Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) (ab150120) at 1/1000 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) at 1/1000 dilution.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (symmetric di methyl R26) antibody [EPR17719] - BSA and Azide free (AB249954)

This data was developed using ab177284, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (Mouse embryonic fibroblast) cells labeling Histone H3 (symmetric di methyl R26) with ab177284 at 1/1000 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 NIH/3T3 cell line.The nuclear counterstain 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 1/1000 dilution (red).

The negative controls are as follows : -

-ve control 1 : ab177284 at 1/1000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) (ab150120) secondary antibody at 1/1000 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 1/1000 dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (symmetric di methyl R26) antibody [EPR17719] - BSA and Azide free (AB249954)

This data was developed using ab177284, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded mouse colon tissue labeling Histone H3 (symmetric di methyl R26) with ab177284 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) 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.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (symmetric di methyl R26) antibody [EPR17719] - BSA and Azide free (AB249954)

This data was developed using ab177284, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded rat colon tissue labeling Histone H3 (symmetric di methyl R26) with ab177284 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear 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

Supplier Data

Western blot - Anti-Histone H3 (symmetric di methyl R26) antibody [EPR17719] - BSA and Azide free (AB249954)

This data was developed using ab177284, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% BSA/TBST.

All lanes:

Western blot - Anti-Histone H3 (symmetric di methyl R26) antibody [EPR17719] (<a href='/en-us/products/primary-antibodies/histone-h3-symmetric-di-methyl-r26-antibody-epr17719-ab177284'>ab177284</a>) at 1/2000 dilution

Lane 1:

NTera-2/D1 (Human malignant pluripotent embryonic carcinoma) whole cell lysate at 10 µg

Lane 2:

HL-60 (Human promyelocytic leukemia cells) whole cell lysate at 10 µg

Lane 3:

HeLa (Human epithelial cells from cervix adenocarcinoma) whole 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: 15 kDa

Observed band size: 15 kDa

false

Exposure time: 30s

• WB

Supplier Data

Western blot - Anti-Histone H3 (symmetric di methyl R26) antibody [EPR17719] - BSA and Azide free (AB249954)

This data was developed using ab177284, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% BSA/TBST.

All lanes:

Western blot - Anti-Histone H3 (symmetric di methyl R26) antibody [EPR17719] (<a href='/en-us/products/primary-antibodies/histone-h3-symmetric-di-methyl-r26-antibody-epr17719-ab177284'>ab177284</a>) at 1/2000 dilution

All lanes:

Human ovary 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: 15 kDa

Observed band size: 15 kDa

false

Exposure time: 10s

• WB

Supplier Data

Western blot - Anti-Histone H3 (symmetric di methyl R26) antibody [EPR17719] - BSA and Azide free (AB249954)

This data was developed using ab177284, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% BSA/TBST.

All lanes:

Western blot - Anti-Histone H3 (symmetric di methyl R26) antibody [EPR17719] (<a href='/en-us/products/primary-antibodies/histone-h3-symmetric-di-methyl-r26-antibody-epr17719-ab177284'>ab177284</a>) at 1/2000 dilution

Lane 1:

F9 (Mouse embryonic carcinoma cell line) whole cell lysate at 10 µg

Lane 2:

D3 (Mouse embryonic stem cell) whole cell lysate at 10 µg

Lane 3:

NIH/3T3 (Mouse embryonic fibroblast cells) whole 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: 15 kDa

Observed band size: 15 kDa

false

Exposure time: 10s

• PepArr

Supplier Data

Peptide Array - Anti-Histone H3 (symmetric di methyl R26) antibody [EPR17719] - BSA and Azide free (AB249954)

This data was developed using ab177284, the same antibody clone in a different buffer formulation.ab177284 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.