Auto-developing Protein Marker Kit for Western Blotting
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Solution Overview
Problem
Current protein markers for Western blotting face challenges such as manual copying from nitrocellulose paper, low accuracy due to heterogeneity and electric charge alterations, and the need for additional labeling which complicates the process, while existing kits use fixed and irregular molecular weights.
Innovation Solution
A regularly-weighted protein molecular weight marker kit comprising recombinant proteins with specific tags (like GST, MBP, and TRX) that auto-develop on film, allowing for accurate recognition by anti-tag antibodies without the need for additional labeling, and can be mixed with solvents for stabilization.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If pre-stain protein markers are used, then the staining results show heterogeneity and electricity alteration, but the molecular weight accuracy is low
Solution Approach 1:
The patent changes the molecular weight parameters of the protein markers to follow a regular arithmetic sequence (5kDa, 10kDa, 15kDa, 20kDa, 25kDa, 30kDa, 35kDa, 40kDa, 45kDa, 50kDa) with uniform intervals, replacing the irregular molecular weights of conventional markers. This parameter standardization directly improves both measurement precision and staining homogeneity by eliminating heterogeneity in the marker proteins themselves.
2Productivity
If manual copying from nitrocellulose paper is performed, then the process is time-consuming, but the automation level is low
Solution Approach 1:
The patent enables the protein markers to self-develop on the film through their inherent properties. The markers contain specific epitopes that can be directly recognized by antibodies conjugated to enzymes, allowing automatic signal generation on the film without requiring manual copying operations. This self-service mechanism dramatically improves productivity while increasing automation extent.
Solution Approach 2:
The patent replaces the mechanical manual copying process with an immunological and enzymatic system. Instead of physically copying markers from nitrocellulose paper, the system uses antibody-antigen recognition and enzyme-substrate reactions to automatically generate visible bands on the film, substituting mechanical operations with biochemical mechanisms.
3Ease of operation
If additional labeling is required for detection, then the procedure becomes troublesome, but the detection capability is enhanced
Solution Approach 1:
The patent designs the protein markers to serve multiple functions simultaneously: they act as molecular weight standards, contain recognizable epitopes for antibody binding, and provide automatic detection capabilities. The markers are constructed with universal epitopic sequences that can be detected by commonly used antibodies, eliminating the need for additional specialized labeling while maintaining enhanced detection capability.
4Manufacturing precision
If fixed and irregular molecular weights are used, then the markers are easier to manufacture, but the regularity and accuracy are compromised
Solution Approach 1:
The patent segments the protein markers into distinct modules with specific molecular weight contributions. Each marker protein is constructed from standardized epitopic sequences combined with proteins of known molecular weights (e.g., GST, MBP, TRX), allowing systematic assembly to achieve regular 5kDa intervals. This modular segmentation enables precise control over molecular weight while maintaining manufacturability through standardized cloning procedures.
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
The kit provides an auto-developing, regularly-weighted protein marker that enhances accuracy and convenience in Western blotting by allowing auto-development on film and stability, overcoming the limitations of existing markers.
Implementation Method 1
accurate recognition by anti-tag antibodies
Implementation Method 2
auto-developing... on film
Implementation Method 3
one or more solvents for stabilizing the recombinant proteins
Data Source
AI summary
The present invention provides an auto-developing and regularly-weighted protein molecular weight marker kit, comprising: (a) a plurality of recombinant proteins having formula (I),(B)m-A-(C)n (I),wherein A is a polypeptide of SEQ ID NO: 1, B and C are independently any mutually identical or different polypeptides with the value of molecular weight being a multiple of 5, and m and n is independently 0 or any integer larger than 0; and (b) one or more solvents for stabilizing the recombinant proteins. The present invention also provides a method for preparing the auto-developing and regularly-weighted protein molecular weight marker kit.