Anti-Histone H3 (acetyl K4) antibody [EPR16596] - BSA and Azide free

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (acetyl K4) antibody [EPR16596] - BSA and Azide free (AB240195)

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 H3 (acetyl K4) with ab176799 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 expression 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 1/1000 dilution (red).

The negative controls are as follows :
-ve control 1 : ab176799 at 1/1000 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 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 1/1000 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab176799).

• ChIP

Unknown

ChIP - Anti-Histone H3 (acetyl K4) antibody [EPR16596] - BSA and Azide free (AB240195)

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 ab176799 (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).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab176799).

• Dot

Lab

Dot Blot - Anti-Histone H3 (acetyl K4) antibody [EPR16596] - BSA and Azide free (AB240195)

Dot blot analysis of Histone H3 (acetyl K4) labeled with ab176799 at 1/1000 dilution. Lane 1 : Human H3(acetyl K4) peptide a Lane 2 : Human H3(methyl K4) peptide c Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) was used as secondary antobody at 1/100000 dilution. Blocking/Dilution buffer : 5% NFDM/TBST. Exposure time : 180 minutes. This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab176799).

• Dot

Supplier Data

Dot Blot - Anti-Histone H3 (acetyl K4) antibody [EPR16596] - BSA and Azide free (AB240195)

Dot blot analysis of Histone H3 (acetyl K4) labeled with ab176799 at 1/1000 dilution.

Lane 1 : H3 (acetyl K4) peptide

Lane 2 : H3 (formylK4) peptide

Lane 3 : H3 (methylK4) peptide

Lane 4 : H3 (crotonylK4) peptide

Lane 5 : Unmodified H3 peptide

Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated was used as secondary antobody at 1/1000 dilution.

Blocking/Dilution buffer : 5% NFDM/TBST.

Exposure time : 3 minutes.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab176799).

• PepArr

Lab

Peptide Array - Anti-Histone H3 (acetyl K4) antibody [EPR16596] - BSA and Azide free (AB240195)

ab176799 (0.1μg/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 was used as the secondary antibody. Protein Array Washing Buffer, 2*10 mins and 2*5 mins. Blocking buffer was 5% BSA in TBST and Innoscan 710 scanner was used. 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. This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab176799).