cross-linking signal with Western Blot

Dongpo Cai dcai at BU.EDU
Sun Oct 12 21:50:16 EST 1997


I am trying to do cross-linking between IL-7 and its receptors using a
recommbinant IL-7
with a Flag epitope as ligand, which has good bioactivity. I also have
the MAb for this
epitope. According to the model developed in the lab I am working, the
epitope is not
located in the ligand-receptor interface, so the MAb should bind to it
after cross-linking.
However I just couldn't get the signal under various conditions I ever
tried. The chemical
linking I used are DSS and BS3. I use human cell line Nalm6, Molt4 which
can express
IL-7R. As far as I know, few people in this world ever did cross-linking
assay with
Western Blot and people prefered to use 125 Iodine which is much more
sensitive. Can
any body give me some advices or suggestions about this problem ? The
reply can also
be sent to dcai at bu.edu. Thanks a lot.




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