Protocol:
1:5000 for immunoblotting
5-7.5 ug for IP (Protein G kit and Co-IP kit)
For immunoblotting I dilute in 5% low fat milk powder in PBS-T and incubate overnight at 4°C.
Dynabeads kits (Invitrogen) were used for IP. Longer incubation of the antibody in the Ab binding and washing buffer of 1 h was used for optimal binding. Bcl-2 has the same size as the light chain, thus crosslinking must be conducted for co-IP
Review: Worked very well overall as it could easily IP Bcl-2 from overexpressing cells and yield a single band by immunoblotting.
On October 29, 2017 Heidi Gytz Olesen, McGill University, Canada wrote:
Mouse anti-Bcl-2 (clone Bcl-2-100), Sigma B3170
Protocol:
1:5000 for immunoblotting
5-7.5 ug for IP (Protein G kit and Co-IP kit)
For immunoblotting I dilute in 5% low fat milk powder in PBS-T and incubate overnight at 4°C.
Dynabeads kits (Invitrogen) were used for IP. Longer incubation of the antibody in the Ab binding and washing buffer of 1 h was used for optimal binding. Bcl-2 has the same size as the light chain, thus crosslinking must be conducted for co-IP
Review: Worked very well overall as it could easily IP Bcl-2 from overexpressing cells and yield a single band by immunoblotting.