The application of OT-2 workstation in protein quantitative determination mainly relies on its highly automated pipetting, mixing, incubation, washing and other functions, as well as its seamless integration with other detection equipment. This all-round automation capability greatly promotes It ensures the efficient execution of the protein quantitative determination process, ensures the accuracy and repeatability of experimental results, and provides convenience for scientific researchers.
1. Automated operation process of OT-2 workstation 1. Sample preparation: Perform necessary preprocessing on the sample to be tested, such as lysis, centrifugation, etc., to obtain the protein solution. 2. Program writing: Use Opentrons software (such as Opentrons App) to write automated experimental programs and set parameters such as pipetting volume, mixing times, and incubation time. 3. Automated pipetting and mixing: The OT-2 platform automatically completes operations such as pipetting of samples, adding and mixing of reagents according to the program, ensuring the accuracy and consistency of each step. 4. Incubation: Place the mixed sample in the temperature control module and incubate according to the set temperature and time to fully combine the antibody with the antigen. 5. Washing: After the incubation, the OT-2 platform automatically performs a washing step to remove unbound reagents and impurities. 6. Signal detection: Transfer the washed sample to a detection instrument (such as a spectrophotometer) to detect the signal intensity after the reaction.
2. Combined with protein quantitative determination methods OT-2 workstation supports a variety of protein quantitative determination methods, such as Coomassie Brilliant Blue method, UV spectrophotometry, BCA method, Folin-phenol reagent method, etc. The chemistry of these methods varies, but all rely on reactions between proteins and specific reagents to generate a detectable signal. 1. Coomassie Brilliant Blue method: After Coomassie Brilliant Blue dye binds to protein, the absorbance at a specific wavelength (such as 595nm) is directly proportional to the protein content. The OT-2 workstation can automatically complete the sample loading, staining, washing and absorbance detection steps. 2. Ultraviolet spectrophotometry: Aromatic amino acids (such as tyrosine and tryptophan) in proteins have characteristic absorption peaks (such as 280nm) under ultraviolet light. The OT-2 workstation can use a UV spectrophotometer to detect the absorbance of the sample at a specific wavelength to calculate the protein content. 3. BCA method: BCA (bicinchoninic acid) reagent reacts with copper ions in the protein under alkaline conditions to form a purple-red complex. The absorbance of the complex is proportional to the protein content. The OT-2 workstation can automatically complete steps such as sample loading, reagent addition, incubation and absorbance detection. 4. Folin-phenol reagent method: Under alkaline conditions, Folin-phenol reagent reacts with tyrosine and tryptophan residues in proteins to generate a dark blue mixture of molybdenum blue and tungsten blue. Its absorbance is directly proportional to the protein content. The OT-2 workstation can also automate various steps of this method.
3. Advantages and features of OT-2 workstation 1. High precision: The OT-2 platform uses high-precision pipettes to ensure the accuracy of the pipetting volume and reduce errors. 2. High efficiency: Automated operation greatly shortens the experimental cycle and improves experimental efficiency. 3. High repeatability: The automated process reduces the uncertainty caused by human operations and improves the repeatability of experimental results. 4. Flexibility: The OT-2 platform can be flexibly configured according to experimental needs and supports a variety of quantitative kits and detection methods. 5. Cost saving: Automated operation reduces the waste of reagents and consumables and reduces experimental costs.
3. Application examples Take the research of the Novo Nordisk Foundation Protein Research Center at the School of Health Sciences at the University of Copenhagen as an example. They used the Opentrons OT-2 automated pipetting platform to design a simple and efficient proteomics sample preparation workflow. , capable of processing up to 192 samples in 6 hours. This workflow integrates sample digestion, purification, and Evotip loading in a fully automated manner to achieve highly sensitive protein quantification.
The application of OT-2 workstation in protein quantitative determination gives full play to its advantages of automation, high precision, high efficiency and high repeatability, providing strong support for biomedical research, drug development, clinical diagnosis and other fields. With the continuous advancement of technology and the in-depth expansion of applications, the OT-2 workstation will demonstrate its unique value and potential in more fields.
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