Culturing Nerve Cells
A do-it-yourself manual for culturing nerve cells, complete with recipes and protocols.
Because neurons and glia in culture are remarkably similar to those in situ, culture systems make it possible to identify significant cell interactions and to elucidate their mechanisms. This book is in many ways a do-it-yourself manual for culturing nerve cells, complete with recipes and protocols. But it also provides an understanding of the principles behind the protocols. In effect the contributors invite you into their labs and provide much of the information you would obtain from such a visit.The authors of the introductory chapters present the nuts-and-bolts principles of growing nerve cells. The authors of the following chapters discuss the culturing of specific cell types. They explain how their experimental goals have shaped their particular cell culture approach and the advantages and disadvantages of the cell culture systems they have developed. They provide detailed protocols and describe their cultures in practical terms, from when the cells are first plated through the various phases of their development.
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immunofluorescence, commonly is used. Antibodies directed against
immunoglobulins from the species used to prepare the cellspecific antibody are
adsorbed to the panning dish. Then the cellspecific antibody may be applied to
the dish as a ...
This can be done by using physiological methods, but often it is more convenient
to immunostain cultures with antibodies directed against the neurotransmitter
itself or against its biosynthetic enzymes. For example, spinal motorneurons have
For most purposes, we prefer immunofluorescence localization (where a labeled
antibody is visualized directly from its emitted fluorescence) rather than
immunocytochemistry (where an antibody coupled to an enzyme is visualized by
Application of Primary Antibody Block in 10% BSA or 10% serum in PBS for 30
min. Incubate in primary antibody, diluted in 2-3% BSA or serum in PBS, for 2h at
room temperature or 37°C or overnight at 4°C. Rinse 3 x 5 min in PBS. Detection
secondary antibody. It does not take into account nonspecific binding of the
primary antibody, which will be detected and amplified by the secondary antibody
. For antibodies generated against a synthetic peptide, the optimal control for ...
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