Hello,
I am working on bovine beta casein which is one of milk proteins. This
bovine beta casein (wild-type) does not have any potential N-linked
glycosylation sites in its stuctural gene. For some reason we introduced
N-glycosylation site (Asn-X-Ser) into bovine beta casein gene and generated
transgenic mice carrying bovine beta casein gene (wild-type or mutant
containing single glycosylation site). We found glycosylated and
unglycosylated bovine beta casein in the milk of transgenic mice using
bovine beta casein specific antibody and confirmed N-linked glycosylation
using PNGase F treatment. However, I have difficulty in detecting
glycosylated beta casein using lectins. The lectin blots with Con A, WGA,
ECA and UEA-1 did not show any positive reactions. So I am wondering what
the reason is. If N-linked oligosaccharides are sulfated, what kind of
method can I try to detect N-linked sugar ?
Thank you.
Byung-Kwon Choi
