CelLytic™ B Cell Lysis Reagent

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	- Cells are lysed by sonication in lysis buffer [400 mM NaCl, 20 mM Tris⋅HCl, pH 8.0, lysozyme 600 μg/mL, 4 μL of Benzonase Dnase (Sigma), 0.5 mL of CelLytic B (Sigma), 1 tablet of EDTA-free protease inhibitor mixture (cOmplete; Roche), 1% (vol/vol) Triton X-100].

Protocol tips
- Cells are lysed by sonication in lysis buffer [400 mM NaCl, 20 mM Tris⋅HCl, pH 8.0, lysozyme 600 μg/mL, 4 μL of Benzonase Dnase (Sigma), 0.5 mL of CelLytic B (Sigma), 1 tablet of EDTA-free protease inhibitor mixture (cOmplete; Roche), 1% (vol/vol) Triton X-100].

Publication protocol

Cells were lysed by sonication in lysis buffer [400 mM NaCl, 20 mM Tris⋅HCl, pH 8.0, lysozyme 600 μg/mL, 4 μL of Benzonase Dnase (Sigma), 0.5 mL of CelLytic B (Sigma), 1 tablet of EDTA-free protease inhibitor mixture (cOmplete; Roche), 1% (vol/vol) Triton X-100]. Previously cleared lysates were subject to ultraspeed centrifugation at 150,000 × g for 1 h at 4 °C to concentrate the sample and remove soluble proteins not assembled as sheaths, as previously reported. In addition, the use of Triton X-100 in lysis buffer reduced the possibility to copurify membrane structures and outer membrane vesicles during the ultracentrifugation.

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

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