Development of a novel protein multi-blotting device

Hagyousif, Amin M.; Chong, Voon Joe; Yokota, Hiroki; Chien, Stanley

doi:10.4236/jbise.2010.312146

Cited by 2 publications

(2 citation statements)

References 5 publications

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“…To elevate efficiency of Western blotting, we have previously developed BlotMan that enables to generate multiple blotting membranes from a single polyacrylamide gel [6] [7]. In a conventional Western blotting procedure, a single gel is used to make a single membrane.…”

Section: Introductionmentioning

confidence: 99%

Improved Protein Transfer Efficiency and Signal Intensity in BlotMan Using Pulse Width Modulation

Truongvo¹,

Zheng²,

Chien³

et al. 2016

JBiSE

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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 anti-tumor 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 Blot-Man reduces an amount of protein samples by generating multiple membranes from a single gel and improving signal intensity with PWM voltage signals.

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Section: Introductionmentioning

confidence: 99%

Improved Protein Transfer Efficiency and Signal Intensity in BlotMan Using Pulse Width Modulation

Truongvo¹,

Zheng²,

Chien³

et al. 2016

JBiSE

Self Cite

View full text Add to dashboard Cite

show abstract

“…Western blotting is a technique widely used to determine the relative amount of proteins in biological samples [1][2][3]. The procedure includes three major steps such as electrophoretic fractionation, transfer (blotting) to a mem-brane, and antibody reactions.…”

Section: Introductionmentioning

confidence: 99%

Voltage profile generation for simultaneous multi-protein detection in western blot analysis

Blair¹,

Wanis²,

Swarnkar³

et al. 2012

JBiSE

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Western blotting is a popular technique for examining expression levels of proteins using gel-based electrophoretic fractionation followed by blotting and antibody reactions. Although this is a mature technique, one of the major limitations is the need to prepare an individual electrophoretic gel for each of the protein species to be analyzed. Since most analyses require the detection of multiple protein species, a procedure that allows utilization of a single gel for detecting multiple protein species should significantly save time and resources. In this paper, we developed a novel multiprotein detection device, which enabled simultaneous detection of several proteins species from a single electrophoretic gel. In this device, a protein transfer unit utilized a multi-anode plate that generated a non-uniform voltage profile. This voltage profile enabled uniform transfer regardless of molecular mass of proteins. <i>In vitro </i> experiments using samples, isolated from boneforming osteoblast cells, showed that the expression levels of 5 - 7 different proteins were detectable in the presence and absence of mechanical stimulation that activated genes necessary for bone formation. The result supports the notion that through simultaneous detection of multiple protein species, the described device contributes to reduction in procedural time and sample amounts, as well as a removal of variations among multiple gels

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Development of a novel protein multi-blotting device

Cited by 2 publications

References 5 publications

Improved Protein Transfer Efficiency and Signal Intensity in BlotMan Using Pulse Width Modulation

Improved Protein Transfer Efficiency and Signal Intensity in BlotMan Using Pulse Width Modulation

Voltage profile generation for simultaneous multi-protein detection in western blot analysis

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