i am working with a protein which is 10kda.i am not able see a band at the expected region in westernblotting-things which i hav used
1.amersham ECL NCM -poresize 2u
2.0.1%SDS in blot buffer
3..63A for 30-45 min
4.0.3% Tween-20 in PBS for wasing and blocking
plz suggest me few things abt were i am going wrong.