Irina Mathov wrote as:
>I'm using biotinilated succinyl wheat germ to develope some western blots
>as Pierce indicates in the instructions but so far I have doubtfull results.
>When I use glycoproteins known to have terminal N Acetyl glucosamine the bands
>are very weak (I have tried different conc. of SWG)
>I couldn't find any references about the needs of cations in the solutions
>used for incubation (as for ConA).
>Does anyone know if SWG needs any cation to improve its binding?
>Any idea will be wellcome.
>Thanks a lot
irina at idehu.edu.ar
I believe WGA also needs cations like Ca2+ and Mg2+. Atleast for affinity
chromatography I normally use PBS with Ca2+, Mg2+ and azide. Also may be
you need to try just biotinylated WGA instead of S-WGA.
I hope this helps.