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[PubMed]5. The presses lyse cells by pressurizing the cell suspension and suddenly releasing the pressure. Of note, this procedure detects some 10% of antigens implying that the real numbers of antigens is considerably higher than detected. Breaking the cell's membranes to retrieve smaller organelles, particles, proteins and nucleic acids is common practice. Immune responses to intracellular bacteria. (A) Outline of the experimental procedure: MelJuSo cells stably transfected with Tap1-GFP 17 were Alexa647-L243 Ab stained for 15min at 4oC before the unbound antibody was removed. [PubMed]8. [PubMed]10. Materials and MethodsFluorescent cell sort stainingMelJuSo (MJS) cells stably transfected with Tap1-GFP were trypsinized, washed and stained with mouse anti human L243-Alexa 647 or L243-Cy5 for 15 minute at 4oC to exclude endocytosis of the MHC-II surface molecules, then washed 3 times to remove traces of antibody.Membrane disruptionThe integrity of the cells was compromised by an EMBL 8.020mm “cell cracker” homogenizer with ball sizes 8.010, 8.008 or 8.004nm (gap of 0.005, 0.006 or 0.008nm between ball and wall of chamber, respectively) either 2, 10 or 30 times.

2001;3(6):E145147. Here, we described that this assumption in not completely correct as novel vesciles are formed by sheering forces. Proteomics of regulated secretory organelles. Flow analysis of MHC molecules and other membrane proteins in isolated phagosomes. [PubMed]4. An alternative lysis method is to freeze the cells directly in liquid nitrogen and ground the frozen cells to a powder using a mortar and pestle that are chilled with liquid nitrogen. The high operating pressures, however, result in a rise in operating temperatures. Jacques Neefjes, The Netherlands Cancer Institute, Division of Cell Biology, Plesmanlaan 121, 1066 CX Amsterdam, The Netherlands.

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Ven 17 Juin - 15:27 (2016)
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