CelLytic™ B Cell Lysis Reagent

Protein isolation Bacteria - Anabaena

Experiment
Protein isolation Bacteria - Anabaena
Product
CelLytic™ B Cell Lysis Reagent from Sigma-Aldrich
Manufacturer
Sigma-Aldrich

Protocol tips

Protocol tips
- Pelleted cells are resuspended in 4 mL of CelLyticTM B containing 10 μg/mL (w/v) lysozyme and shaken on a vortexer for 30 min. After, samples are centrifuged (10,000g, RT, 5 min).

Publication protocol

The cell lysis assay was conducted by pelleting 7 mL of cells that had been grown in BG-11/HEPES medium under WL at 15 μmol m-2 s-1 as described above and diluted to an OD750 of 0.4 and incubated for an additional 24 h. Pelleted cells were then resuspended in 4 mL of CelLyticTM B (Millipore Sigma, St. Louis, MO, United States) containing 10 μg/mL (w/v) lysozyme and shaken on a vortexer for 30 min. After this period, samples were centrifuged at 10,000 ×g at room temperature for 5 min. The level of cellular lysis was estimated based on measuring absorbance at 660 nm of the chlorophyll released into the supernatant. Chlorophyll absorbance values were standardized relative to total soluble protein content of cells.

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Discussion

Discussion

2 years ago

Author: Hollie Fowler United Kingdom

How can I deal with my pellet being too viscous?

I am using 8M Urea to lyse my cells but after centrifugation my pellet is very viscous. This causes me trouble when I try to collect the supernatant. Is there any way to avoid it and do you think it will have an effect on the concentration of my protein?

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Papers

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Manufacturer protocol

Download the product protocol from Sigma-Aldrich for CelLytic™ B Cell Lysis Reagent below.

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