Short protocol:
1:200 for immunoblotting
IP: 2 μg (Protein G kit)/6.4 μg (co-IP kit)
For immunoblotting, I dilute in 5% low fat milk powder in PBS-T and incubate overnight at 4°C.
For IP, Dynabeads kits (Invitrogen) were used according to manufacturers recommendations.
Review:
Works well for detection of the VDAC1 protein. VDAC1 has the same size as the light chain, thus crosslinking to Dynabeads must be performed. We were unable to capture VDAC1 with Dynabeads® Co-Immunoprecipitation Kit (Invitrogen), however the protein G kit version may have worked (no crosslinking beforehand)
On June 10, 2020 Jeannette Nardelli, Université de Paris, NeuroDiderot wrote:
Works great for WB in a dilution of 1:1000.
Works roughly for IF on frozen sections.
For more information see Journiac et al. Cell Reports 2020, “ Cell Metabolic Alterations due to Mcph1 Mutation in Microcephaly”.
https://www.sciencedirect.com/science/article/pii/S221112472030396X
On October 29, 2017 Heidi Gytz Olesen, McGill University, Canada wrote:
Rabbit anti-VDAC1, Abcam Ab15895
Short protocol:
1:200 for immunoblotting
IP: 2 μg (Protein G kit)/6.4 μg (co-IP kit)
For immunoblotting, I dilute in 5% low fat milk powder in PBS-T and incubate overnight at 4°C.
For IP, Dynabeads kits (Invitrogen) were used according to manufacturers recommendations.
Review:
Works well for detection of the VDAC1 protein. VDAC1 has the same size as the light chain, thus crosslinking to Dynabeads must be performed. We were unable to capture VDAC1 with Dynabeads® Co-Immunoprecipitation Kit (Invitrogen), however the protein G kit version may have worked (no crosslinking beforehand)