Ready-to-use Solution for Protein Extraction from E. coli / Yeast Cells

　1 is a ready-to-use solution for protein extraction from cultured Escherichia coli (E. coli) / yeast cells. By suspending cells in 1 and then centrifuging, the supernatant containing proteins can be obtained. Extracted protein can be used in downstream applications such as electrophoresis and western blotting.

Application example

[Protein extraction]

1. Centrifuge E. coli at 5000 × g and yeast at 3000 × g for 10 minutes.
2. Remove as much medium as possible from the pellet.
3. Resuspend pellet in 1. Use 2-4 mL buffer per gram of pellet and mix at room temperature for 10 minutes.
4. Centrifuge E. coli at 15000 × g and yeast at 14000 × g for 10 minutes and collect lysate fraction.

[Protein quantification]: measurement of protein concentration in lysate fraction by pyrogallol red method

[Electrophoresis / Western blotting]: Detection of specific protein in lysate fraction

RecA in E. coli (left, 38 kDa) and Rad51 in S. cerevisiae (right, 43 kDa) can be extracted efficiently.

References

• 1) Non-detergent sulphobetaines: a new class of mild solubilization agents for protein purification
• L. Vuillard, C. Braun-Breton, T. Rabilloud, Biochem. J. 1995, 305, 337.
• 2) A Highly Efficient Procedure for the Extraction of Soluble Proteins from Bacterial Cells with Mild Chaotropic Solutions
• V. N. Danilevich, L. E. Petrovskaya, E. V. Grishin, Chem. Eng. Technol. 2008, 31, 904.

Related Products

P2575

Pyrogallol Red (Ready-to-use solution) [for Protein determination]

D5909

DAB staining kit

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