Validated for:

• Western blot-WB
• Immunofluorescence-IF
• Immunoprecipitation, the reagent of choice for this application-IP
• Chromatin immunoprecipitation-ChIP

Anti-acetyl lysine antibody (AAC03) was raised against a proprietary mix of acetylated proteins to ensure broad acetyl lysine recognition. It detects a wide range of acetylated proteins, including tubulins and histones. AAC03 is the antibody of choice for IP applications as it gives the richest acetylated protein IP profile. AAC03 is purified via Protein G chromatography.

Host/isotype/clone:

• Mouse/IgG1
• Clone: 19C4B2.1

Reactivity:

• All species (acetyl-lysine is identical in all species)

Amount of material:

• AAC03: 2 x 100 µl when reconstituted (100 – 150 µg per tube)
• AAC03-S: 1 x 25 µl when reconstituted (25 – 37.5 µg per tube)

Working concentrations:

• WB 1:1000 dilution 1-1.5 µg/ml final 200 ml working volume - full size Ab
• IF 1:4000 dilution 1-1.5 µg/ml final 800 ml working volume - full size Ab
• IP 20 ul neat 20 µg per IP 10 reactions – full size Ab
• ChIP 1:100 dilution 10-15 µg/ml final 20 ml working volume – full size Ab

Formulation

• Supplied as a lyophilized powder
• Buffer composition after reconstitution in 50% glycerol in water: 200 mM PIPES pH7.0, 5% sucrose, 1% dextran, 50% glycerol

Sensitivity:

• Detects as little as 20 pg acetylated BSA in WB application

Specificity:

• High selectivity: Chemically acetylated BSA (50 pg) is detected in WB, while 1 µg of non-acetylated BSA is not—demonstrating a ≥20,000-fold specificity.
• Enhanced signal with HDAC inhibition: Treatment with Trichostatin A (TSA), an HDAC inhibitor, results in >20-fold increase in signal for acetylated proteins in IP assays and > 10-fold for WB.
• IF selectivity: Cells treated with HDAC inhibitors show strong enhancement of acetylation signals in the nucleus and along microtubules, consistent with known acetylation targets such as histones and tubulin.