Short protocol:
1:2000 for immunoblotting
1.4 μg/1.5mg beads for IP (Dynabeads® Protein G kit)
For immunoblotting I diluted in 5% low fat milk powder in PBS-T and incubate overnight at 4oC.
Dynabeads kits (Invitrogen) for IP were used according to manufacturers recommendations.
Review: The antibody efficiently precipitated endogenous Opa1 from Hek293 cells, though excessive co-elution of the immunoglobulins were detected as well. Several unspecific bands were present upon immunoblotting, however the Opa1 protein was clearly noticeable.
On October 29, 2017 Heidi Gytz Olesen, McGill University, Canada wrote:
Rabbit anti-Opa1, Abcam Ab42364
Short protocol:
1:2000 for immunoblotting
1.4 μg/1.5mg beads for IP (Dynabeads® Protein G kit)
For immunoblotting I diluted in 5% low fat milk powder in PBS-T and incubate overnight at 4oC.
Dynabeads kits (Invitrogen) for IP were used according to manufacturers recommendations.
Review: The antibody efficiently precipitated endogenous Opa1 from Hek293 cells, though excessive co-elution of the immunoglobulins were detected as well. Several unspecific bands were present upon immunoblotting, however the Opa1 protein was clearly noticeable.