Hi,

Can anyone suggest a method to crosslink the Antibody to the sepharose beads ?? (an option apart from using commercially available kits)..

thank u

arun

here is a method I saw on the web
Crosslinking Ab PDF

I've never tried it personally, but worth a look

btw this is a stupid question but crosslinked antibodies are not supposed to come down with the elute
by heat/boiling right?
my PI told me he thinks it probably will...but...I'm really confused now!!
(told me not to try the method )

thank u lynn... I will look into that.

but I didn't understand why you said its a stupid question..

if you don't crosslink the Ab with the beads using some chemical, the Ab will come out from the beads when u heat it in the sample buffer .. right ??

lynn.. the problem is that my protein of interest is around 50kDa

hence if I don't crosslink the Ab to the beads, the heavy chain fragments are going to interfere in the western..