A simple monolithic column electroelution for protein recovery from gel electrophoresis.
Protein recovery from gel electrophoresis plays an important role in functional genomics and proteomics but faces a series of issues (e.g., complex procedure, low recovery, long experimental time). In this study, a monolithic column electroelution (MCE) was developed for protein recovery from gel electrophoresis. With the model proteins of bovine serum albumin (BSA), hemoglobin (Hb), and myoglobin (Mb), the developed device and method were compared with common electroelution procedures in agarose gel electrophoresis (AGE). The comparative experiments revealed that (i) the protein recovery achieved with the developed device was greater than 83%, much higher than the 41% to 50% achieved with the common devices; (ii) the running time to obtain 70% recovery was approximately 15 min, evidently shorter than the 240 min with the common devices; and (iii) the device and procedure were simple and less time-consuming as compared with those of the common devices. It was observed that the serum protein bands cut from polyacrylamide gel electrophoresis could be transferred into solution in 15 to 30 min with 82% yield. The device, along with its relevant procedure, has potential use in protein extraction and proteomics as well as in DNA studies.
Laboratory of Analytical Biochemistry and Bioseparation, State Key Laboratory of Microbial Metabolism, School of Life Science and Biotechnology, Shanghai Jiao Tong University, Shanghai, China.
Guo-Qing
GQ
Li
Jing
J
Shao
Chen-Gang
CG
Guo
Jing-Yu
JY
Dong
Liu-Yin
LY
Fan
Cheng-Xi
CX
Cao
D000818:Animals; D002021:Buffers; D002417:Cattle; D004587:Electrophoresis, Agar Gel / Q000295:instrumentation / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D011506:Proteins / Q000032:analysis / Q000302:isolation & purification*; D040901:Proteomics; D012709:Serum Albumin / Q000032:analysis / Q000302:isolation & purification; D013997:Time Factors
D002021:Buffers; D011506:Proteins; D012709:Serum Albumin