Preparation and analysis of total cellular protein samples

Summary

This experiment is based on "Color Atlas of Practical Flow Cytometry", edited by Shukui Wang and Zhenying Zhou.

Operation method

Preparation and analysis of total cellular protein samples

Move

Take 20 mg of fluorescein isothiocyanate, add 20 ml of anhydrous ethanol to fully dissolve FITC, that is, the stock solution of the staining solution, and keep it in the refrigerator at 4℃ for use. The specific fluorescence staining steps are as follows:

1. Dilution of FITC storage solution: Dilute 50 ug/ml working solution with PBS solution of pH 7.4, and set aside.

2. Stained cells were firstly membrane-broken with 70% alcohol for 10 min, so that the FITC dye can enter into the cells.

3. Wash off the membrane-breaking agent by centrifugation with PBS solution, and then centrifugate to collect the cells.

4. Then take 2 ml of FITC working solution and add it into the sample. 4. Add 2 ml of FITC working solution to the sample, mix well, and place in the refrigerator at 4℃ for 30 min.

5. Centrifuge with PBS at 1500 r/min for 5 min to remove the staining solution, and then centrifuge to remove the supernatant.

6. Add 0.5 ml of PBS, mix well, and run the test on the machine.


For more product details, please visit Aladdin Scientific website.

https://www.aladdinsci.com/

Categories: Protocols

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