Quantitative method for dimensional electrophoresis protein sample

Abstract

The invention discloses a quantitative method for a dimensional electrophoresis protein sample. The method includes the steps of: preparing a dimensional electrophoresis sample; preparing a 1-2mg / mL bovine serum albumin (BSA) solution by using the a dimensional electrophoresis cell lysate, taking 8 centrifuge tubes, adding 0-200 muL of bovine serum albumin solution respectively, and supplementing ultrapure water to 1 mL to reach a final concentration of standard protein of 0-0.2mg / ml, then adding 5 mL of a 0.01% Coomassie brilliant blue G-250 or R-250 dye solution into each centrifuge tube, and fully mixing to obtain standard mixture solutions with different concentrations; preparing a dimensional electrophoresis sample mixed solution; adding the sample decrypting and framing fluorescence intensity; drawing a standard curve; and calculating the concentration of the dimensional electrophoresis protein sample. The invention has the advantages of simple operation, accuracy, fastness and cheapness.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a protein quantification method. [0002] Background technique [0003] Protein quantification is an important link in cell and molecular biology experiments. Commonly used protein quantification methods include the biquinolinic acid method (BCA method) and the Coomassie brilliant blue method (Bradford method) invented by Bradford MM in 1976, both of which are quantitative methods based on absorbance values. Two-dimensional electrophoresis (2-D) is the core technology in the process of proteomics research. Due to the complex process of 2-D protein sample processing, a wide variety of reagents are used. Reagents such as urea, thiourea, Triton X-100 (Triton X-100), sodium dodecyl sulfate (SDS) and other reagents used in this process are easy to react with BCA and Coomassie brilliant blue (CBB) to develop color, thereby These two methods cannot be used in 2-D experiments because they a...

AI Technical Summary

Problems solved by technology

Reagents such as urea, thiourea, Triton X-100 (Triton X-100), sodium dodecyl sulfate (SDS) and other reagents used in this process are easy to react with BCA and Coomassie brilliant blue (CBB) to develop color, thereby Affects absorbance values ​​and thus quantification accuracy, so neither method can be used for 2-D experiments

In view of the above reagent compatibility problems, Bio-Rad (Bio-Rad) and General Electric (GE) have developed protein quantification kits specifically for 2-D, but these kits are developed based on spectrophotometry , there are many operation steps, high requirements for the experimenter, and the kit is expensive