MicroRotofor™ Cell Lysis Kits

MicroRotofor lysis kits provide cell lysis and protein extraction protocols that are tailored to the specific needs of different sample sources. Convenient and easy to use, all four kits are based on the same chaotropic protein solubilization buffer (PSB), which contains nondetergent sulfobetaine 201 (NDSB 201) along with urea, thiourea, and CHAPS for particularly effective solubilization (Vuillard et al. 1995). The kits generate total protein samples that are ready for fractionation with the MicroRotofor liquid-phase IEF cell, but they may also be applied to other downstream applications, such as SDS-PAGE and 2-D gel electrophoresis.

Species-Specific Protocols
Different sample types have different requirements for effective cell disruption, and all four kits combine PSB with other elements to accommodate these specific needs.

• MicroRotofor cell lysis kit (mammal) — uses maceration and solubilization (up to a 100 mg sample) using ReadyPrep mini grinders and PSB (Vuillard et al. 1995). The kit is intended for use with samples from mammalian sources, but it may also be applied to other animal tissues
• MicroRotofor cell lysis kit (bacteria) — for use with both gram-negative and gram-positive bacterial cultures. It employs enzymatic digestion using lysozyme followed by solubilization into PSB (Vuillard et al. 1995)
• MicroRotofor cell lysis kit (plant) — for use with soft plant tissues and cultured cells. It employs maceration (up to a 1 g sample) and solubilization into PSB. A ReadyPrep 2-D cleanup kit is provided to enable removal of plant-derived phenolics, which may interfere with IEF
• MicroRotofor cell lysis kit (yeast) — for use with yeast cultures (~60 µl wet cell pellet). It employs enzymatic digestion of the cell wall with lyticase followed by solubilization into PSB (Scott et al. 1980, Vuillard et al. 1995)

References
Scott J et al. (1980). Lyticase: endogluconase and protease activities that act together in cell lysis. J Bacteriol 142, 414–423.
Vuillard L et al. (1995). Non-detergent sulphobetaines: a new class of mild solubilization agents for protein purification. Biochem J 305, 337–343.