Specificity of GA3512. Western
blotting was performed with cell lysates from HEK293 cells transfected
to express hBest1, hBest2, hBest3, mBest1-EGFP, mBest2, or mBest3.
Untransfected HEK-293 cells were included as a negative control. A
hBest2 and mBest2, but was more
efficient at identifying hBest2. It did not crossreact with other
: A commercially available anti-Best2 antibody
obtained from Novus Biologicals (NOVUS) was also specific to hbest2 and
: B4947A, a rabbit polyclonal antibody against mBest2
strongly recognized mBest2 and was less effective at identifying
hBest2. Controls confirm expression of D
: hBest1, E
mBest1-EGFP, and F
: mBest3 using the antibodies indicated.
Interestingly, only anti-Best2 antibodies recognized both the human and
mouse forms. Also, note the approximately 70 kDa band that is
present in every lane in E
. This band is non-specific [1
equivalent loading, blots were cut and the bottom portion blotted for
human β-actin (hb-actin).