Acetyl-Lysine Affinity Beads - AAC03

Acetyl-Lysine Affinity Beads

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AAC03-Beads

Cytoskeleton offers two types of Acetyl-Lysine Affinity Beads, Cat # AAC02-Beads and Cat # AAC03-Beads. Both reagents are comprised of mouse anti-acetyl-lysine antibodies that are covalently linked to protein G beads and both enrich a broad range of acetylated proteins. AAC02-Beads and AAC03-Beads can be combined to give a more extensive acetylated protein enrichment profile (Cat # AAC04-Beads is a 1:1 combination of AAC02-Beads:AAC03-Beads).

AAC03-Beads has an overlapping but unique specificity profile when compared to AAC02-Beads and may outperform AAC02-Beads when examining a specific target protein, see Detailed methods and Validated Applications for examples.  As acetylation generally represents a very small percent of total target protein, utilizing optimized reagents is extremely critical to success. When examining the acetylation state of a new protein of interest (POI), it is recommended to try AAC03-Beads and AAC02-Beads separately to determine which reagent is optimal for your POI.

Each lot of affinity-bead is quality controlled to provide high batch to batch consistency, see COA documents.

Validated Applications

Application 1: Detection of Acetylation Changes in a Target POI

AAC03-Beads have been shown to be a superior reagent for the immunoprecipitation of acetylated EGFR and RhoGDI Target POI (Fig. 1). In the presence of HDAC inhibitors AAC03-Bead IPs show a an 11.3 fold and a 2.5 fold enhanced acetylation of EGFR and RhoGDI respectively.  While AAC02-Bead shows a weaker 8 fold and 1.1 fold increase respectively.

Figure 1 Legend: AAC03-Beads & AAC02-Beads (50µl bead slurry) and a 1:1 mix (25 µl each) AAC04-Beads, were used to IP acetylated proteins from Cos-7 cells either treated (+) or untreated (-) with deacetylase inhibitors, [TSA (1µM) and nicotinamide (1mM)], for 6 hours. Western blot analysis using anti EGFR and anti -RhoGDI antibodies was performed and signals were quantitated using LiCor Empiria software. IP assays were also carried out and signals quantitated for mouse IgG control beads (blots not shown). Each IP used 1 mg of lysate.

 

 

aac04_fig_2

Application 2: Immunoprecipitation of Total Acetyl-Lysine Profile

AAC03-Beads show a robust total acetyl IP profile with an overlapping but unique specificity profile to AAC02-Beads.  AAC03-Beads can be used alone or in combination with AAC02-Beads to study the acetylome.

Figure 2 Legend: AAC02-Beads, AAC03-Beads, and AAC04-Beads (50µl bead slurry) were used to IP acetylated proteins from Cos-7 cells either treated (+) or untreated (-) with deacetylase inhibitors [TSA (1µM) and nicotinamide (1mM)] for 6 hours. The total profile of enriched acetylated proteins were eluted and analyzed by western blot with an AAC03-HRP antibody (1:3000).  Mouse IgG beads are used as a control for non-specific binding (Cat # CIG02). Each IP assay utilized 1 mg of Cos-7 lysate

 

 

aac04_fig1_2

Amount:
Each package contains enough acetyl-lysine affinity beads for 40 reactions.

 

For more information contact: signalseeker@cytoskeleton.com

Associated Products:

Signal-Seeker™ SUMOylation 2/3 Detection Kit (Cat. # BK162)

Signal-Seeker™ Ubiquitination Detection Kit (Cat. # BK161)

Signal-Seeker™ Acetyl-Lysine Detection Kit (Cat. # BK163)

Signal-Seeker™: BlastR™ Rapid Lysate Prep Kit (Cat. # BLR01)

Signal-Seeker™: PTMtrue™ Aceetyl lysine Antibody (Cat.# AAC03)

For product Datasheets and MSDSs please click on the PDF links below.   For additional information, click on the FAQs tab above or contact our Technical Support department at tservice@cytoskeleton.com

AuthorTitleJournalYearArticle Link
He, Ying et al.PPARγ Acetylation Orchestrates Adipose Plasticity and Metabolic RhythmsAdvanced Science2023ISSN 2198--3844
Oldfield C. et. al.Muscle-specific sirtuin 3 overexpression does not attenuate the pathological effects of high-fat/high-sucrose feeding but does enhance cardiac SERCA2a activityPhysiol Rep.2021ISSN 3440-5591
Kumar, Manish et al.Inhibition of histone acetyltransferase function radiosensitizes CREBBP/EP300 mutants via repression of homologous recombination, potentially targeting a gain of functionNature Communications2021ISSN 2041-1723
Horita, Henrick et al.Utilizing a comprehensive immunoprecipitation enrichment system to identify an endogenous post-translational modification profile for target proteinsJournal of Visualized Experiments2018ISSN 1940-087X
Horita, Henrick et al.Utilizing optimized tools to investigate PTM crosstalk: Identifying potential PTM crosstalk of acetylated mitochondrial proteinsProteomes2018ISSN 2227-7382

If you have any questions concerning this product, please contact our Technical Service department at tservice@cytoskeleton.com