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Titlebook: Immunocytochemical Methods and Protocols; Lorette C. Javois Book 19992nd edition Humana Press 1999 DNA.ELISA.Filtration.Organelle.PCR.cell

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Overview of Antibody Use in Immunocytochemistry the antigenic sites (direct method) (.) (. Chapter 15). Later, the more sensitive and versatile indirect method was introduced (.) (. Chapters 16–18). In this method, the specific antibody, bound to the antigen, was detected with a secondary reagent, usually another antibody that had been tagged wi
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Purification of Antibodies Using Ammonium Sulfate Fractionation or Gel Filtration charge, the surface distribution of polar side chains, and the size of the polypeptide, as well as the pH and temperature of the solution. Immunoglobulins precipitate at 40–50% ammonium sulfate saturation depending somewhat on the species and subclass (.). The desired saturation is brought about ei
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Purification of Antibodies Using Protein A-Sepharose and FPLCfor isolating immune complexes or immunoglobulins from a crude solution (.,.). Protein A has also been useful for separating Fc fragments from Fab fragments after proteolytic di gestion. The advantages of protein A are mainly its stability and specificity. Protein A is stable over a wide pH range (p
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Processing of Cytological Specimens a dilute medium are best suited for the preparation of cytospins through cytocentrifugation. Cell suspensions that exist in a high-viscosity medium, are best suited to be tested as swab preparations (.). The constant among these preparations is that the whole cell is present on the slide surface. F
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Processing of Tissue Specimensmple immersion fixation. It should be mentioned, though, that depositing large organs like whole brains in buckets of fixative may provide fine cellular detail but will probably result in the loss of some labile brain proteins desired for study (.). The individual who actually obtains the specimen i
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Direct Immunofluorescent Labeling of Cellsfluorescence is weak since only one labeled primary antibody binds to each antigen. The indirect labeling technique adds one more incubation and wash step, but it has the advantage of amplifying the fluorescent signal because several fluorescently labeled secondary antibodies can bind to each primar
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Fluorescence Labeling of Surface Antigens of Attached or Suspended Tissue-Culture Cellshighly useful diagnostic tools in clinical medicine. The identification of surface antigenic molecules using antibodies can be performed by several methods, including immunofluorescence microscopy, enzyme and discrete marker microscopy, enzyme and radioactive labeling of mass cultures, and flow cyto
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