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Hi everyone,
 
I have a question regarding the capture of fluorescent proteins on agarose
beads.

I¹ve captured four different fluorescent proteins on agarose beads by two
different methods and have obtained different images, but was expecting very
similar results.
 
Method 1 ­ Fluorescent protein was captured on agarose beads using an
antibody (against the fluorescent protein) and results show bright
expression with a Ohalo effect¹ of fluorescence around the agarose beads.
 
Method 2 ­ Fluorescent proteins containing the S-tag sequence (at the C
terminal of protein) were captured on agarose beads coated with S-protein
(S-tag binds to S-protein) and the images obtained were not the Ohalo
effect¹ as seen previously but the bead outline and fluorescence expression
appear fuzzy.
 
Has anyone come across this before or have any ideas as to what could be
causing the differences seen?
 
Any help would be greatly appreciated.
 
Nisha