The acetylated lysine antibody is immobilized to agarose beads via amide linkages. The product can be utilized as an affinity matrix for rapid isolation and purification of proteins/peptides with acetyl lysine residues.
The acetylated peptides eluted from acetyl-lysine-specific affinity chromatography (ICP0388).
Approximately 5 mg of trypsinated crude protein from TSA treated MMRU cells wereloaded.
The maximum binding of acetylated BSA with ICP0388-2MG.
-50 µl of ICP0388-2MG 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)
Formulation
0.5 mL of agarose beads suspended in 1 mL of slurry
Antibody Immobilized
4 mg/ml antibody is covalently linked through amide bonds with NHS activated-SMCC, which further conjugates to thiolated agarose beads via thiol ether bonds.
Specificity
The antibody selectively captures proteins/peptides wtih acetylated lysine residues (N-epsilon). No cross reactions to methylated proteins/peptides.
BindingCapacity
Approximately 0.2 mg of acetylated histone / mL slurry.
Applications
Rapid isolation and purification of proteins/peptides with acetylated lysine residues from cell lysate or protease-digested mixtures.
Scientific Description
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.
Storage & Stability
Product is stable for 30 days at room temperature. For extended storage, store product at -20°C. Shake thoroughly before use.Expiration date is one year from date of shipping if properly stored.