Anti-Methylated Lysine Western Blot

1.

After the protein transfer, block the mebrane with 3% BSA (do not use milk products, as they may contain competitive mehtylated proteins) in PBST at room temperatrue for 60 min.

2.

Wash the membrane with PBSt once and incubate with anti-methylated lysine antibody (0.25-1 μg/mL in 0.1%BSA-PBST) at room temperature for 120 min.

3.

Wash the membrane with PBST four times and incubate with anti-rabbit IgG HRP (0.1-0.25 μg/mL in 0.1%BSA-PBST) at room temperature for 60 min.

4.

Wash the membrane with PBST four times and add ECL solution for 2 min then expose to the film.