2 replies to this topic

[mt2]

Hi,

I have a query. I was trying to reduce background/non specific bands I was seeing through pre-blocking my agrose a beads with 5% BSA prior to incubating with antibody and lysate for a co-ip. However this lead to a greatly increased background. Why would this be?

Regards,

M

[bob1]

Antibody binds to something in the BSA?

[mt2]

I incubated my agarose beads in 5% BSA before washing twice in my lysis buffer. I then added antibody and lysate and proceeded. Surely BSA should bind to any non specific binding beads. Why therefore would this affect antibody binding and lead to increased background? Sorry if this is basic.