Experimental determination of protein concentration by spotted filter membrane binding method

Summary

The spotted filter binding method is a rapid method for the initial determination of protein content. This method is particularly useful for screening useful fractions in density gradient centrifugation or column chromatography. It can also be used to indicate the presence of protein peaks to reduce the number of samples for further protein quantification. Source: Handbook of Protein Technology

Operation method

spotted filter membrane binding method

Materials and Instruments

Protein samples to be tested
10% trichloroacetic acid (TCA), Coomassie blue gel staining solution, Coomassie blue gel decolorizing solution.
Whatman 3 MM filter paper

Move

1. use a pencil to draw 1 cm x 1 cm squares on 3 mm filter paper;

2. Add 3 ul of sample to the center of each square. 3;

3. Allow the filter paper to dry for about 15 minutes. 4;

4. immerse the filter paper in 10% TCA solution for 30 s (TCA solution can be reused);

5. Rinse the filter paper quickly with water. 6;

6. immerse the filter paper in a Caullemont blue gel stain for 30 s (the stain is reusable). 7. rinse the filter paper quickly with water;

7. rinse the filter paper quickly with water. 8;

8. decolorize the filter paper in a Caulmers blue gel decolorizing solution, decolorization is completed within a few minutes;

9. the presence of a certain amount of protein can be determined by the corresponding blue squares.

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