Development of a novel protein multi-blotting device
Amin M. Hagyousif, Voon J. Chong, Hiroki Yokota, Stanley Y. P. Chien
DOI: 10.4236/jbise.2010.312146   PDF    HTML     4,289 Downloads   7,795 Views   Citations


Blotting is a common technique widely used for molecular analysis in life sciences. The Western blot, in particular, is a process of transferring protein samples from a polyacrylamide gel to a blotting membrane and detecting the levels of specific proteins through reactions with primary and secondary antibodies. The state-of-the-art of Western blotting usually generates one blotting membrane per gel. However, multiple copies of blots are useful in many applications. Two blotting copies from a single protein gel, for instance, can be used for identifying a total amount of proteins of interest as well as its specific subpopulation level such as a phosphorylated isoform. To achieve this multi-blotting operation from a single gel, we modified a blotting procedure and developed a novel blotting device. The device consisted of a multi-anode plate and a microcontroller. It was designed to generate a well-controlled electrophoretic voltage profile, which allowed a quasi-uniform transfer of proteins of any size. The prototype device was built and its operation procedure was described. The experimental results clearly supported the notion that the described device was able to achieve multiple blotting from a single gel and reduce time and cost for protein analysis.

Share and Cite:

Hagyousif, A. , Chong, V. , Yokota, H. and Chien, S. (2010) Development of a novel protein multi-blotting device. Journal of Biomedical Science and Engineering, 3, 1125-1132. doi: 10.4236/jbise.2010.312146.

Conflicts of Interest

The authors declare no conflicts of interest.


[1] Towbin, H., Staehelin, T. and Gordon, J. (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets procedure and some applications. Proceedings of the National Academy of Sciences (USA), 76, 4350-4354.
[2] Chen, H. (2003) Semi-dry electroblotter. US Patent 6,592,734 B2.
[3] Adcock, M. (1990) Field inversion elecroblotting & electroelution. US Patent 4959133.
[4] Zahringer, H. (2008) Product survey: Blotting equipment, low-tech at its best. Lab Times, May, 55-59.
[5] Ajit, V. (2009) Essentials of glycobiology. 2nd Edition, Cold Spring Harbor Laboratories Press.
[6] Tarrant, M.K. and Cole, P.A. (2009) The chemical biology of protein phosphorylation. Annual Review Biochemistry, 78, 797-825.
[7] Quercioli, V. (1993) Pulse width modulated (PWM) power supplies, Vol. 45(Studies in Electrical and Electronic Engineering). Elsevier, Amsterdam.
[8] Wang, D., Christensen, K., Chawla, K., Xiao, G., Krebsbach, P.H., and Franceschi, R.T. (1999) Isolation and characterization of MC3T3-E1 preosteoblastsubclones with distinct in vitro and in vivo differentiation/miner- alization potential. Journal of Bone Mineral Research, 14, 893-903.
[9] Hirasawa, H., Jiang, C., Zhang, P., Yang, F.C. and Yokota, H. (2010) Mechanical stimulation suppresses phosphorylation of eIF2?and PERK-mediated responses to stress to the endoplasmic reticulum. FEBS Letters, 584, 745-752.
[10] Hamamura, K. and Yokota, H. (2007) Stress to endoplasmic reticulum of mouse osteoblasts induces apoptosis and transcriptional activation for bone remodeling. FEBS Letters, 581, 1769-1774.

Copyright © 2024 by authors and Scientific Research Publishing Inc.

Creative Commons License

This work and the related PDF file are licensed under a Creative Commons Attribution 4.0 International License.