where are they on SDS-PAGE gel of Co-IPed complex??? - (May/10/2011 )
Assume protein Tom (MW 50) and Jack (MW 80) form complex in my cells. I used anti-Tom antibody to co-IP protein Jack(pull down Jack）. Assume I did successfully pull down Jack. My question is after boiling my protein A beads with 2xSample buffer and loaded to SDS-PAGE, where is Jack located on the gel? Is it at position of MW 80, because it aready dissociated with Tom? or MW 130, if they are still in a complex? Thank you!
-fortunate-
fortunate on Tue May 10 23:55:11 2011 said:
Assume protein Tom (MW 50) and Jack (MW 80) form complex in my cells. I used anti-Tom antibody to co-IP protein Jack(pull down Jack）. Assume I did successfully pull down Jack. My question is after boiling my protein A beads with 2xSample buffer and loaded to SDS-PAGE, where is Jack located on the gel? Is it at position of MW 80, because it aready dissociated with Tom? or MW 130, if they are still in a complex? Thank you!
Hola, you have to search Jack in its position at 80Kd because with the denaturation for loading samples the comples will be broken. If you see the band at 130 use a new prepared loading buffer. Buena suerte
-protolder-