In vitro binding assay
purpose
to determine in vitro binding of M9 and transportin.
Protocol
1.pull-down (PBS-Tran /S100T + GST-A1/GST-M9, methylation or not)
2.western---- loading ¡÷ running ¡÷ transfer to membrane
3.blotting---- block ¡÷ 1 antibody ¡÷ 2 antibody ¡÷ ECC ¡÷ film ¡÷ stain
result
in vitro binding of GST-A1 (or GST-M9) and transportin is negative.