Convenient analysis of protein modification by chemical blotting with fluorogenic "click" reagents.
Direct visualization of bioorthogonal alkyne or azide handles using fluorogenic azide-alkyne cycloaddition conducted on the surface of a blot membrane. The method eliminates the need for separation steps to remove excess small molecule reagents before attachment of antigen molecules or other visualization handles, and is especially useful for the analysis of peptides and small proteins. A variety of potential fluorogenic reagents are assessed, and sensitivity (<0.1 picomole) similar to current commercially available fluorescence imaging methods is possible.
Department of Chemistry, Rice University, MS 60, 6100 Main Street Houston, Texas 77005, USA. zb1@rice.edu.
Jun
J
Ohata
Farrukh
F
Vohidov
Zachary T
ZT
Ball
D000480:Alkynes / Q000737:chemistry; D000818:Animals; D001386:Azides / Q000737:chemistry; D002417:Cattle; D057930:Click Chemistry / Q000379:methods*; D005456:Fluorescent Dyes / Q000737:chemistry / Q000378:metabolism*; D010455:Peptides / Q000737:chemistry / Q000378:metabolism; D011499:Protein Processing, Post-Translational; D012710:Serum Albumin, Bovine / Q000378:metabolism
D000480:Alkynes; D001386:Azides; D005456:Fluorescent Dyes; D010455:Peptides; D012710:Serum Albumin, Bovine