I also tried Anti-BirA (Abcam’s ab14002) polyclonal Chicken western (1:1000) using HEK cell lysates overexpressing a BirA fusion construct (After Transmembrane domain). observed Unspecific bands around 129, 75, 50 and 45 kDa including Control Hek lysate (Un-transfected)
Used Abcam’s ab14002 polyclonal Chicken-anti-BirA on WB using HEK cell lysates overexpressing a BirA fusion construct, but I can’t recommend that. Lot’s of a-specific staining. I also saw that Abcam discontinued this antibody, I guess they came to the same conclusion.
I’ve used Abcam’s ab14002 polyclonal Chicken-anti-BirA at 5.5 µg/ml in blockbuffer (1%BSA in PBS) on transiently transfected, 4% PFA fixed, 1% triton X100 permeabilized HEK293 and RN22 cells (type 1 transmembrane fusion protein w/ the BirA just extracellular from the membrane). Secondary was LifeTech’s Goat-anti-Chicken Alexa 488. Primary staining o/n at 4C, secondary 1 h at RT.
The staining suffered high a-specific background in HEK cells and less but definitely still present background in the RN22 cells. It was clearly possible to distinguish specific staining from background, but I was not too impressed with the staining in general.
Didn’t test it in any other modality, concentration, or protocol yet.
On November 30, 2015 Arulmani Manavalan wrote:
I also tried Anti-BirA (Abcam’s ab14002) polyclonal Chicken western (1:1000) using HEK cell lysates overexpressing a BirA fusion construct (After Transmembrane domain). observed Unspecific bands around 129, 75, 50 and 45 kDa including Control Hek lysate (Un-transfected)
On November 30, 2015 Maarten Rotman wrote:
Used Abcam’s ab14002 polyclonal Chicken-anti-BirA on WB using HEK cell lysates overexpressing a BirA fusion construct, but I can’t recommend that. Lot’s of a-specific staining. I also saw that Abcam discontinued this antibody, I guess they came to the same conclusion.
On June 29, 2015 Maarten Rotman, Mayo clinic, Jacksonville wrote:
I’ve used Abcam’s ab14002 polyclonal Chicken-anti-BirA at 5.5 µg/ml in blockbuffer (1%BSA in PBS) on transiently transfected, 4% PFA fixed, 1% triton X100 permeabilized HEK293 and RN22 cells (type 1 transmembrane fusion protein w/ the BirA just extracellular from the membrane). Secondary was LifeTech’s Goat-anti-Chicken Alexa 488. Primary staining o/n at 4C, secondary 1 h at RT.
The staining suffered high a-specific background in HEK cells and less but definitely still present background in the RN22 cells. It was clearly possible to distinguish specific staining from background, but I was not too impressed with the staining in general.
Didn’t test it in any other modality, concentration, or protocol yet.