Conductive Polymer Gel Transfer Unit for Western Blotting
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Solution Overview
Problem
Current western blotting techniques face challenges such as air bubble introduction, gel tearing, and the need for separate apparatuses for protein separation and transfer, which complicates the process and limits visualization during electrophoresis.
Innovation Solution
A single precast gel and membrane combination unit using conductive polymers that allows for both protein separation and transfer in a single step, with transparent conductive plates enabling visualization during electrophoresis and perpendicular current flow for efficient protein transfer to a blotting membrane.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Device complexity
If traditional separate apparatuses are used for protein separation and transfer, then each step can be performed with dedicated equipment, but the process complexity increases and requires multiple apparatuses
Solution Approach 1:
The patent combines protein separation and transfer functions into a single apparatus. The gel supporting plate serves dual purposes: it provides structural support during electrophoresis and acts as a transfer electrode during protein transfer. This merging eliminates the need for separate apparatuses and reduces process complexity while maintaining both functions.
Solution Approach 2:
The gel supporting plate is designed with universal functionality to perform multiple roles. It serves as both the structural support for the gel during separation and as the transfer electrode for protein transfer. This multi-functionality reduces the number of components needed and simplifies the overall system.
2Illumination intensity
If transparent materials are used for gel supporting plates, then visualization of protein separation is enabled, but electrical conductivity for protein transfer is reduced
Solution Approach 1:
The gel supporting plate is constructed from composite materials that combine transparency and electrical conductivity. Specifically, it uses transparent conductive polymers or transparent conductive coatings on transparent substrates. This composite structure allows simultaneous achievement of visualization capability through transparency and electrical conductivity for efficient protein transfer.
3Quantity of substance
If gels are made thinner to reduce protein requirements, then less protein is needed for analysis, but gel tearing occurs during transfer
Solution Approach 1:
The patent merges the gel supporting plate with the transfer electrode function. During protein transfer, electrical current is applied through the gel supporting plate itself rather than requiring physical manipulation of the thin gel. This eliminates mechanical stress on thin gels and prevents tearing while enabling transfer of gels containing minimal protein amounts.
4Reliability
If conductive blotting electrodes are used during separation phase, then protein transfer can be enabled, but the electric field is nullified and protein migration is prevented
Solution Approach 1:
The system dynamically switches between separation and transfer modes by controlling electrode activation. During the separation phase, only the separation electrodes are activated while the transfer electrodes (including the gel supporting plate) remain inactive. During the transfer phase, the separation electrodes are deactivated and the transfer electrodes are activated. This dynamic control prevents electric field interference and allows both functions to be performed effectively.
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
This approach simplifies the western blotting process by allowing hands-free protein separation and efficient transfer without gel reorientation, providing a fast and reliable method for protein analysis.
Implementation Method 1
The gel is capable of separating proteins by size within the gel when an electric current flows between electrodes on opposite sides of the y-axis of the gel
Implementation Method 2
the current flows through a conductive plate or semi-conductive plate in a direction perpendicular to the direction of the flow of the current during the separation phase
Data Source
AI summary
A precast gel and membrane combination unit for use in gel electrophoresis and membrane transfer for protein analysis, and method of use. Transparent electrically conductive polymer plate(s) house an electrophoresis gel and immunoblotting membrane. The gel and membrane are positioned between two plates of conductive polymers or plates having an electrically conductive layer or film. During the electrophoresis step, electric current moves proteins through the gel allowing for protein separation. Then, without removing or reorienting the gel or apparatus, the electrical contacts are switched to allow the flow of electricity to run perpendicularly through the gel via the conductive polymers, which will allow the proteins to transfer to the membrane housed in the same precast gel and membrane combination unit. The apparatus allows the user to visualize the steps of protein separation and protein transfer without transferring the gel between electrophoresis and transfer steps.