CelLytic™ B Cell Lysis Reagent

Protein isolation Bacteria - Bacillus cellulosilyticus

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

Protocol tips

Protocol tips
- Before adding CelLytic B reagent resuspend bacterial pellets in 1.0 ml of 50 mM Tris-HCl, pH 9.5 (three times).

- The pellets are lysed by shaking at 37°C for 30 minutes in 1.0 ml of CelLytic B reagent in 50 mM Tris-HCl, pH 9.5.

- The pellets are recovered by centrifugation and resuspended in 1.0 ml of 50 mM Tris-HCl, pH 9.5 (three times). Finally the pellets are resuspended in 1.0 ml of 50 mM Tris-HCl, pH 9.5.

Publication protocol

Cultures were harvested at 24 hours by centrifugation. Pellets were resuspended in 1.0 ml of 50 mM Tris-HCl, pH 9.5 and pelleted; this was repeated three times. The pellets were then lysed by shaking at 37°C for 30 minutes in 1.0 ml of CelLytic B reagent in 50 mM Tris-HCl, pH 9.5. The pellets containing the cell walls were recovered by centrifugation and resuspended in 1.0 ml of 50 mM Tris-HCl, pH 9.5 and pelleted; this was repeated three times. Finally the pellets were resuspended in 1.0 ml of 50 mM Tris-HCl, pH 9.5.

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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

Check out relevant papers found by Labettor's AI that are relevant for performing Protein isolation Bacteria - Bacillus cellulosilyticus using CelLytic™ B Cell Lysis Reagent from Sigma-Aldrich.

Paper title
Genomic and Enzymatic Results Show Uses a Novel Set of LPXTA Carbohydrases to Hydrolyze Polysaccharides
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Manufacturer protocol

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

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