Validated for:

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

Anti-acetyl lysine antibody (AAC02) 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, and the mitochondrial protein pyruvate dehydrogenase (PDHE1). AAC02 shows some specificity towards acetylated histones and mitochondrial proteins in IF applications. It is purified via Protein G chromatography.

Host/isotype/clone:

• Mouse/IgG2b
• Clone: 7B5A1

Reactivity:

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

Amount of material:

• AAC02: 2 x 100 µl when reconstituted (100 – 150 µg per tube)
• AAC02-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 1:1000 dilution 1-1.5 µg/ml final 200 ml working volume - full size Ab
• IP 20 ul neat 20-30 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:

• 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.
• IF selectivity: Cells treated with HDAC inhibitors show strong enhancement of acetylation signals in the nucleus, along microtubules, and in mitochondria, consistent with known acetylation targets such as histones, tubulin, and PDHE1.