Labeling lipids for imaging in live cells.
Cold Spring Harb Protoc
; 2010(7): pdb.prot5459, 2010 Jul 01.
Article
em En
| MEDLINE
| ID: mdl-20647363
ABSTRACT
INTRODUCTION:
Fluorescently tagged lipid-binding domains have become a popular tool to image lipids that are involved in intracellular signaling processes. The readout usually involves the translocation of the lipid-binding domain from the cytosol or nucleosol to the membrane of interest, or vice versa. Unfortunately, this method seems to work predominantly for lipids in the plasma membrane, whereas lipids such as phosphatidylinositol 4,5-bisphosphate (PIP(2)) are not recognized in the membranes of the endoplasmic reticulum or the Golgi. Very recently, we developed an alternative way of localizing a lipid of interest by fluorescent labeling of minimally modified lipid derivatives using a single specific chemical reaction. For lipid location analyses, the method is used in fixed cells. However, for studying lipid dynamics, specific labeling in living cells is also possible. This protocol describes how to directly label lipids for imaging in living cells.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Coloração e Rotulagem
/
Imageamento Tridimensional
/
Lipídeos
Limite:
Humans
Idioma:
En
Ano de publicação:
2010
Tipo de documento:
Article