Cd44 high background

Protein Detection Protein Labels
1

Hi, I am using BD’s CD44-PE (IM7 clone) at 1:200 (recommended 1:50) with PBS/2% FBS 1mM EDTA and appear to have a high background with MCF7 cells. Ab incubation 15min on ice.

Anybody have suggestions?

Thanks

2

Hi Sachin

That may be attributed to Hight Ab concentration as well as non specific binding to the membrane. You can try to increase the dilution of Ab and may be try upto 1:1000 or even more if the binding gives sufficient signal without background. Alternatively, you can Increase the duration of blocking.