Signal corresponding to Bak was weaker than aspecific signal. Specificity was confirmed on Bak KO cells. Dilution 1:1000 in 1% milk in PBS-T
For more information see Rizzotto et al. Cell Reports 2020, “Nutlin-Induced Apoptosis Is Specified by a Translation Program Regulated by PCBP2 and DHX30”. https://doi.org/10.1016/j.celrep.2020.03.011
Rabbit anti-Bak (G-23), Santa Cruz Biotechnology, Sc-832.
1:500 for immunoblotting
2 μg/1.5mg beads for IP (Dynabeads® Protein G kit)
Short protocol:
For immunoblotting I dilute in 5% low fat milk powder in PBS-T and incubate overnight at 4°C.
Dynabeads kits (Invitrogen) were used according to manufacturers recommendations.
Review:
Unspecific bands were present after immunoblotting, however the Bak protein itself was clearly visible as well. We were able to immuno precipitate endogenous Bak from HEK293 cells, though the background from the Bak antibody was very high.
On April 22, 2020 Dario Rizzotto wrote:
Signal corresponding to Bak was weaker than aspecific signal. Specificity was confirmed on Bak KO cells. Dilution 1:1000 in 1% milk in PBS-T
For more information see Rizzotto et al. Cell Reports 2020, “Nutlin-Induced Apoptosis Is Specified by a Translation Program Regulated by PCBP2 and DHX30”. https://doi.org/10.1016/j.celrep.2020.03.011
On October 29, 2017 admin wrote:
This antibody has been discontinued
On October 29, 2017 Heidi Gytz Olesen, McGill University, Canada wrote:
Rabbit anti-Bak (G-23), Santa Cruz Biotechnology, Sc-832.
1:500 for immunoblotting
2 μg/1.5mg beads for IP (Dynabeads® Protein G kit)
Short protocol:
For immunoblotting I dilute in 5% low fat milk powder in PBS-T and incubate overnight at 4°C.
Dynabeads kits (Invitrogen) were used according to manufacturers recommendations.
Review:
Unspecific bands were present after immunoblotting, however the Bak protein itself was clearly visible as well. We were able to immuno precipitate endogenous Bak from HEK293 cells, though the background from the Bak antibody was very high.