All lanes : Anti-Histone H3 (acetyl K27) antibody [EPR21237] (ab241152) at 1 µg/ml

Lane 1 : HeLa Whole Cell Lysate
Lane 2 : HeLa Histone Preparation Nuclear Lysate
Lane 3 : NIH3T3 Histone Preparation Nuclear Lysate
Lane 4 : Purified Core Histones from Sodium Butyrate Treated HeLa Cells
Lane 5 : Purified Core Histones from Untreated HeLa Cells

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : HRP conjugated Goat Anti-Rabbit IgG (H+L) at 1/5000 dilution

Developed using the ECL technique.

Performed under reducing conditions.

Predicted band size: 15 kDa


Exposure time: 2 seconds

This blot was produced using a 4-20% Criterion TGX gel in 1X SDS buffer. The gel was run at 180V for 40 minutes before being transferred onto a Nitrocellulose membrane at 25V for 7 minutes using the Trans-Blot turbo transfer system. The membrane was then blocked for an hour using 3% milk before being incubated with ab241152 overnight at 4°C. Antibody binding was detected using a Goat anti-Rabbit antibody conjugated to HRP, and visualised using Bio-Rad Clarity Western ECL substrate.

All lanes : Anti-Histone H3 (acetyl K27) antibody [EPR21237] (ab241152) at 1 µg/ml

Lane 1 : HeLa whole cell lysate
Lane 2 : HeLa triton nuclear lysate
Lane 3 : NIH3T3 triton nuclear lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : HRP conjugated Goat Anti-Rabbit IgG (H+L) at 1/5000 dilution

Developed using the ECL technique.

Performed under reducing conditions.

Predicted band size: 15 kDa


Exposure time: 1 second

This blot was produced using a 4-20% Criterion TGX gel in 1X SDS buffer. The gel was run at 180V for 40 minutes before being transferred onto a Nitrocellulose membrane at 25V for 7 minutes using the Trans-Blot turbo transfer system. The membrane was then blocked for an hour using 3% milk before being incubated with ab241152 overnight at 4°C. Antibody binding was detected using a goat anti-Rabbit antibody conjugated to HRP, and visualised using Bio-Rad Clarity Western ECL substrate.

All batches of ab241152 are tested in Peptide Array against peptides to different Histone H3 and H4 modifications. Six dilutions of each peptide are printed on to the Peptide Array in triplicate and results are averaged before being plotted on to a graph. Results show strong binding to Histone H3 - acetyl K27 peptide (ab24404) indicating that this antibody specifically recognises the Histone H3 - acetyl K27 modification.

ab24404 - H3 K27ac
ab179063 - H2A.Z K11ac
ab14997 - ScH2A K21ac
ab179089 - H2B K20cr
ab179088 - H2B K20ac
ab179246 - H3 K23pr
ab178670 - H2A.Z unmod(1-30)
ab16635 - H3 K9ac
ab179334 - H4 K12cr
ab48359 - H3 K23ac
ab179327 - H4 K8cr
ab179253 - H3 K27cr
ab179030 - H2A R77me1
ab179053 - H2A.X unmod(1-30)