Anti-Histone H4 (acetyl K20) antibody [EPR16998(2)] - ChIP Grade - BSA and Azide free

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Histone H4 (acetyl K20) antibody [EPR16998(2)] - ChIP Grade - BSA and Azide free (AB249948)

This data was developed using ab177191, 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 (acetyl K20) with ab177191 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

The expression increased on HeLa cells after treatment with Trichostatin A (500 ng/ml) for 4 hours.

The nuclear counter stain is DAPI (blue).

Tubulin is detected with Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution and 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 : ab177191 at 1/500 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 (acetyl K20) antibody [EPR16998(2)] - ChIP Grade - BSA and Azide free (AB249948)

This data was developed using ab177191, 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 cell line) cells labeling Histone H4 (acetyl K20) with ab177191 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

The expression increased on NIH/3T3 cells after treatment with Trichostatin A (500 ng/ml) for 4 hours.

The nuclear counter stain is DAPI (blue).

Tubulin is detected with Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution and 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 : ab177191 at 1/500 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.

• WB

Supplier Data

Western blot - Anti-Histone H4 (acetyl K20) antibody [EPR16998(2)] - ChIP Grade - BSA and Azide free (AB249948)

This data was developed using ab177191, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

5 μg peptide is used in each lane of the blocking experiment.

All lanes:

Western blot - Anti-Histone H4 (acetyl K20) antibody [EPR16998(2)] - ChIP Grade (<a href='/en-us/products/primary-antibodies/histone-h4-acetyl-k20-antibody-epr169982-chip-grade-ab177191'>ab177191</a>) at 1/1000 dilution

Lane 1:

HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate treated with 500 ng/ml TSA for 4 hours at 10 µg

Lane 2:

HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate treated with 500 ng/ml TSA for 4 hours at 10 µg with H4K20ac peptide

Lane 3:

HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate treated with 500 ng/ml TSA for 4 hours at 10 µg with H4K20 unmodified peptide

Lane 4:

HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate treated with 500 ng/ml TSA for 4 hours at 10 µg with H4K16Me1 peptide

Lane 5:

HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate treated with 500 ng/ml TSA for 4 hours at 10 µg with H4K16ac peptide

Lane 6:

HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate treated with 500 ng/ml TSA for 4 hours at 10 µg with H4K16 unmodified peptide

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

false

Exposure time: 5s

• WB

Supplier Data

Western blot - Anti-Histone H4 (acetyl K20) antibody [EPR16998(2)] - ChIP Grade - BSA and Azide free (AB249948)

This data was developed using ab177191, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Histone H4 (acetyl K20) antibody [EPR16998(2)] - ChIP Grade (<a href='/en-us/products/primary-antibodies/histone-h4-acetyl-k20-antibody-epr169982-chip-grade-ab177191'>ab177191</a>) at 1/1000 dilution

Lane 1:

Untreated HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 10 µg

Lane 2:

HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate treated with 500 ng/ml TSA for 4 hours 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

false

Exposure time: 30s

• WB

Supplier Data

Western blot - Anti-Histone H4 (acetyl K20) antibody [EPR16998(2)] - ChIP Grade - BSA and Azide free (AB249948)

This data was developed using ab177191, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Histone H4 (acetyl K20) antibody [EPR16998(2)] - ChIP Grade (<a href='/en-us/products/primary-antibodies/histone-h4-acetyl-k20-antibody-epr169982-chip-grade-ab177191'>ab177191</a>) at 1000 Cells

Lane 1:

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

Lane 2:

NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate treated with 500 ng/ml TSA for 4 hours 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

false

Exposure time: 30s

• PepArr

Lab

Peptide Array - Anti-Histone H4 (acetyl K20) antibody [EPR16998(2)] - ChIP Grade - BSA and Azide free (AB249948)

This data was developed using ab177191, the same antibody clone in a different buffer formulation. ab177191 at 0.1 μg/mL 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). A Goat Anti-Rabbit IgG, (H+L), Fluo 647nm conjugated at 1/50000 dilution was used as the secondary antibody. Washing buffer and time was protein Array Washing Buffer, 2*10mins and 2*5mins and blocking buffer was 5% BSA in TBST. 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 the area under the curve when antibody binding values are plotted against the corresponding peptide concentration. Each circle area is normalized to the peptide with the strongest affinity. Scanner details : Innoscan 710