Anti-Histone H4 (acetyl K12) antibody [EPR17906] - BSA and Azide free

• WB

Supplier Data

Western blot - Anti-Histone H4 (acetyl K12) antibody [EPR17906] - BSA and Azide free (AB249998)

This data was developed using ab177793, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Histone H4 (acetyl K12) antibody [EPR17906] (<a href='/en-us/products/primary-antibodies/histone-h4-acetyl-k12-antibody-epr17906-ab177793'>ab177793</a>) at 1/10000 dilution

Lane 1:

HeLa (Human epithelial cells from cervix adenocarcinoma) treated with 500 ng/ml Trichostatin A for 4 hours whole cell lysates at 10 µg

Lane 2:

HeLa (Human epithelial cells from cervix adenocarcinoma) untreated whole cell lysates at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 11 kDa

Observed band size: 11 kDa

false

Exposure time: 15s

• WB

Supplier Data

Western blot - Anti-Histone H4 (acetyl K12) antibody [EPR17906] - BSA and Azide free (AB249998)

This data was developed using ab177793, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Histone H4 (acetyl K12) antibody [EPR17906] (<a href='/en-us/products/primary-antibodies/histone-h4-acetyl-k12-antibody-epr17906-ab177793'>ab177793</a>) at 1/10000 dilution

Lane 1:

NIH/3T3 (Mouse embyro fibroblast cells) treated with 500 ng/ml Trichostatin A for 4 hours whole cell lysates at 10 µg

Lane 2:

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

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 11 kDa

false

Exposure time: 15s

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H4 (acetyl K12) antibody [EPR17906] - BSA and Azide free (AB249998)

This data was developed using ab177793, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Rat colon tissue labeling Histone H4 (acetyl K12) with ab177793 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500. Nucleus staining on Rat colon tissue is observed. Counter stained with Hematoxylin.

Negative control : Using PBS instead of primary antibody.

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 H4 (acetyl K12) antibody [EPR17906] - BSA and Azide free (AB249998)

This data was developed using ab177793, the same antibody clone in a different buffer formulation.

Intracellular Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) treated (Red)/untreated (Green) with 500ng/ml Trichostatin A for 4 hours with purified ab177793 at 1/80 dilution. The secondary antibody was Goat anti rabbit IgG (Alexa Fluor^® 488) at 1/2000 dilution. A Rabbit monoclonal IgG (Black) was used as the isotype control and cells without incubation with primary antibody and secondary antibody (Blue) were used as unlabeled control.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Histone H4 (acetyl K12) antibody [EPR17906] - BSA and Azide free (AB249998)

This data was developed using ab177793, 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, treated with Trichostatin A (500 ng/ml) for 4 hours or untreated, labeling Histone H4 (acetyl K12) with ab177793 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 is observed. Acetylation level increased after treatment with Trichostatin A (500 ng/ml) for 4 hours. 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;
1. ab177793 at 1/2000 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
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 H4 (acetyl K12) antibody [EPR17906] - BSA and Azide free (AB249998)

This data was developed using ab177793, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling Histone H4 (acetyl K12) with ab177793 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500. Nucleus staining on Human colon tissue is observed. Counter stained with Hematoxylin.

Negative control : Using PBS instead of primary antibody.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• PepArr

Unknown

Peptide Array - Anti-Histone H4 (acetyl K12) antibody [EPR17906] - BSA and Azide free (AB249998)

This data was developed using ab177793, the same antibody clone in a different buffer formulation.

ab177793 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.