All lanes : Anti-Histone H3 (acetyl K9) antibody [EPR21239] (ab241151) 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 ab241151 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 K9) antibody [EPR21239] (ab241151) at 1 µg/ml

Lane 1 : HeLa triton nuclear lysate
Lane 2 : NIH3T3 triton nuclear lysate

Lysates/proteins at 10 µg per lane.

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

Developed using the ECL technique.

Performed under reducing conditions.

Predicted band size: 15 kDa


Exposure time: 5 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 ab241151 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 ab241151 are tested in Peptide Array against peptides to different Histone H2, 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 K9 peptide (ab16635), indicating that this antibody specifically recognises the Histone H3 - acetyl K9 modification.

ab16635 - H3 K9ac
ab7768 - H3 K4me2
ab179053 - H2A.X unmod(1-30)
ab178987 - H1.4 unmod(1-30)
ab179205 - H3 R8me1
ab178994 - H2A unmod(1-30)
ab179136 - H3 unmod(1-33)
ab178667 - ScH2A unmod(1-30)
ab179097 - H2B unmod(26-56)
ab179083 - H2B K15me1
ab179106 - H2B K46me1
ab179108 - H2B K46me3
ab179264 - H3 P38hpro
ab179247 - H3 unmod(20-51)