Anti-Histone H4 (crotonyl K8) antibody [EPR17905(R)] - BSA and Azide free

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Histone H4 (crotonyl K8) antibody [EPR17905(R)] - BSA and Azide free (AB251336)

This data was developed using ab201075, 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 H4 (crotonyl K8) with ab201075 at 1/1000 dilution, followed by Goat Anti-rRbbit 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 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 : ab201075 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.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Histone H4 (crotonyl K8) antibody [EPR17905(R)] - BSA and Azide free (AB251336)

This data was developed using ab201075, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Neuro-2a (Mouse neuroblastoma cell line) cells labeling Histone H4 (crotonyl K8) with ab201075 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 Neuro-2a 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 : ab201075 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 H4 (crotonyl K8) antibody [EPR17905(R)] - BSA and Azide free (AB251336)

This data was developed using ab201075, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded mouse colon tissue labeling Histone H4 (crotonyl K8) with ab201075 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear staining on mouse colon 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 H4 (crotonyl K8) antibody [EPR17905(R)] - BSA and Azide free (AB251336)

This data was developed using ab201075, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded rat liver tissue labeling Histone H4 (crotonyl K8) with ab201075 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear staining on rat liver 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 H4 (crotonyl K8) antibody [EPR17905(R)] - BSA and Azide free (AB251336)

This data was developed using ab201075, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% BSA/TBST.

All lanes:

Western blot - Anti-Histone H4 (crotonyl K8) antibody [EPR17905(R)] (<a href='/en-us/products/primary-antibodies/histone-h4-crotonyl-k8-antibody-epr17905r-ab201075'>ab201075</a>) at 1/1000 dilution

Lane 1:

NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate at 10 µg

Lane 2:

HeLa (Human epithelial cell line 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 1/100000 dilution

Predicted band size: 11 kDa

Observed band size: 11 kDa

true

Exposure time: 30s

• WB

Supplier Data

Western blot - Anti-Histone H4 (crotonyl K8) antibody [EPR17905(R)] - BSA and Azide free (AB251336)

This data was developed using ab201075, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% BSA/TBST.

All lanes:

Western blot - Anti-Histone H4 (crotonyl K8) antibody [EPR17905(R)] (<a href='/en-us/products/primary-antibodies/histone-h4-crotonyl-k8-antibody-epr17905r-ab201075'>ab201075</a>) at 1/1000 dilution

Lane 1:

C6 (Rat glial tumor cell line) whole cell lysate at 10 µg

Lane 2:

PC-12 (Rat adrenal gland pheochromocytoma cell line) 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 1/100000 dilution

Predicted band size: 11 kDa

Observed band size: 11 kDa

true

Exposure time: 30s

• PepArr

Unknown

Peptide Array - Anti-Histone H4 (crotonyl K8) antibody [EPR17905(R)] - BSA and Azide free (AB251336)

This data was developed using ab201075, the same antibody clone in a different buffer formulation.

ab201075 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.