Validated for:

• Western blot-WB
• Immunofluorescence-IF
• Immunoprecipitation-IP
• Chromatin immunoprecipitation-ChIP

Anti-acetyl lysine antibody (AAC01) was raised against a proprietary mix of acetylated proteins to ensure broad acetyl lysine recognition. It detects a wide range of acetylated proteins, including tubulin, histones, glutamic dehydrogenase, actins, and Arp 2/3 complex proteins. AAC01 is purified via Protein G chromatography.

Host/isotype/clone:

• Mouse/IgG2b
• Clone: 3C6.08.20

Reactivity:

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

Amount of material:

• AAC01: 2 x 100 µl when reconstituted (100-180 µg per tube)
• AAC01-S: 1 X 25 µl when reconstituted (25-45 µg per tube)

Working concentrations:

• WB 1:1000 dilution 1-1.8 µg/ml final 200 ml working volume - full size Ab
• IF 1:200 dilution 5-9 µg/ml final 40 ml working volume - full size Ab
• IP 20 ul neat 20-36 µg per IP 10 reactions – full size Ab
• ChIP 1:100 dilution 10-18 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: 250 mM PIPES pH 7.0, 1% sucrose, 0.5% dextran, 50% glycerol.

Sensitivity:

• Detects as little as 5 pg acetylated BSA in WB application

Specificity

• High selectivity: Chemically acetylated BSA (5 pg) is detected in WB, while 1 µg of non-acetylated BSA is not—demonstrating a ≥200,000-fold specificity.
• Enhanced signal with HDAC inhibition: Treatment with Trichostatin A (TSA), an HDAC inhibitor, results in >10-fold increase in signal for acetylated proteins in WB and IP assays.
• Reduced signal in the absence of HDAC inhibitors: Lysates prepared without HDAC inhibitors show a greatly diminished signal in both WB and IP.
• IF selectivity: Cells treated with HDAC inhibitors show strong enhancement of acetylation signals in the nucleus and along microtubules, consistent with known acetylation targets like histones and tubulin.