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文献和实验and the supernatant discarded) twice with Solution A (1.2 M sorbitol, 50 mM KPO4, pH 7). To spheroplast, cells are resupended in 500 ul of Solution A containing 0.1 b-mercaptoethanol, 0.02 glusulase and 5ug/ml zymolyase. The suspension is incubated at 37oC
Anti-HA Immunocytology The multiple copies of the HA epitope present in the HAT tag can be detected by the mouse monoclonal antibodies 12CA5 (Boehringer) and 16B12 (MMS101R; BAbCO, Richmond, California) (see Table 1). On Western blots
Isolation of murine splenocytes
listed are for use with one mouse. Supplies 15ml conical tube 60mm petri dish 5ml pipet 100μm cell strainer (can substitute autoclaved fine nylon mesh for Protocol B) 3mL sterile disposable syringe, no needle
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