Author(s)

Thuc Nhi Truongvo¹, Feixue Zheng², Stanley Chien^2,3, Hiroki Yokota^1,2*

¹Department of Biomedical Engineering, Indiana University Purdue University Indianapolis, Indianapolis, IN, USA.
²YC Bioelectric LLC, Indianapolis, IN, USA.
³Department of Electrical and Computer Engineering, Indiana University Purdue University Indianapolis, Indianapolis, IN, USA.

BlotMan is a protein blotting device that allows generating multiple membranes from a single polyacrylamide gel. To transfer all proteins uniformly with the same efficiency regardless of protein size, BlotMan employs pulse-width-modulated (PWM) voltage that applies a higher average voltage to a larger protein species. BlotMan can be controlled not only by its custom-made interface but also by a smart phone via Bluetooth technology. In this study, we examined effects of PWM signals (50%, 60%, and 80% duty cycle) on transfer efficiency and signal intensity in comparison to a constant voltage signal (100% duty cycle). The result revealed that in response to the same average voltage of 150 V, a lower duty cycle with a higher maximum voltage increased transfer efficiency as well as sharpness of transferred proteins. We validated BlotMan’s capability using a chondrosarcoma cell line (SW1353 cells) and a breast cancer cell line (MDA-MB231 cells) in response to antitumor chemical agents. BlotMan successfully generated 5 membranes from a single gel and detected 5 protein species such as c-Src, eukaryotic translation initiation factor 2 alpha (eIF2), phosphorylated eIF2, lamin B, and actin. Collectively, we demonstrated herein that BlotMan reduces an amount of protein samples by generating multiple membranes from a single gel and improving signal intensity with PWM voltage signals.

Pulse Width Modulation (PWM), Western Blotting, Protein Transfer

Truongvo, T. , Zheng, F. , Chien, S. and Yokota, H. (2016) Improved Protein Transfer Efficiency and Signal Intensity in BlotMan Using Pulse Width Modulation. Journal of Biomedical Science and Engineering, 9, 269-279. doi: 10.4236/jbise.2016.95020.