Capturing SDS-treated biotinylated protein and peptide by avidin functional affinity electrophoresis with or without SDS in the gel running buffer.
Avidin functional affinity electrophoresis (AFAEP) is a new method of affinity electrophoresis. In this technique, bifunctional linker glutaraldehyde is added to the polyacrylamide gel solution to embed avidin within the gel matrix by interaction with the amino/amide groups. Samples are heated with triglycine sodium dodecyl sulfate (SDS) sample buffer to ensure that biotinylated proteins biotinylated peptides are negatively charged and migrate electrophoretically toward the cathode through the avidin zone regardless of their pI values. The AFAEP method allows the capture and concentration of biotinylated proteins or biotinylated peptides irrespective of the use of SDS in both the sample buffer and the gel running buffer.
Lee
Bao-Shiang
BS
Protein Research Laboratory, Research Resources Center, University of Illinois, Chicago, IL 60612, USA. boblee@uic.edu
Gupta
Shalini
S
Krishnanchettiar
Sangeeth
S
Lateef
Syed Salman
SS
D000818:Animals; D001360:Avidin / Q000378:metabolism*; D019921:Biotinylation; D002021:Buffers; D002417:Cattle; D002645:Chickens; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D006801:Humans; D010455:Peptides / Q000737:chemistry* / Q000378:metabolism*; D011506:Proteins / Q000737:chemistry* / Q000378:metabolism*; D012967:Sodium Dodecyl Sulfate / Q000494:pharmacology*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D002021:Buffers; D010455:Peptides; D011506:Proteins; D001360:Avidin; D012967:Sodium Dodecyl Sulfate