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The acetylated protein signal as determinated by ELISA titration (OD50 titer).
The acetylated proteins were eluted from 1 mL of Rabbit anti-Acetyl Lysine Agarose (ICP0388) and immobilized on a microplate, then followed by detection with Mouse anti-Acetyl Lysine HRP. Rabbit anti-Biotin Agarose (1 mL) was utilized as the negative control.
The maximum binding of acetylated BSA with ICP0388-5MG.
-50 µl of ICP0388-5MG were incubated with 1 mg of acetylate BSA in a 1 ml tube for 60 min.
– After washing with PBST 4 times, the bound acetylated BSA was eluted with 1 ml of 0.5 M HCl
-5 µl / lane of the eluted acetylated BSA was resolved by SDS-PAGE and blotted wih monoclonal Mouse anti-Acetylated Lysine (ICP0390)
-ECL exposure 3 seconds
Protein acetylation is a form of post-translational modification known to regulate many diverse biological processes. Detection, isolation and identification of acetylated proteins/peptides is essential in proteomic studies. Affinity chromatography is one of the most efficient and rapid methods to enrich and purify the acetylated species for further MS/MS identification.
10 mg/mL antibody is covalently linked through amide bonds with NHS activated-SMCC, which further conjugates to thiolated agarose beads via thiol ether bonds.
Product is stable for 30 days at room temperature. For extended storage, store product at -20°C. Do not aliquot and shake thoroughly before use.Expiration date is one year from date of shipping if properly stored.
