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Metabolic Labeling of Glycoproteins 下载为PDFGeneral Information
This protocol details the preparation of cell lysates metabolically labeled with azido- or alkynylsugar. However, investigators may wish to determine the optimal concentration of metabolic
reagent as well as labeling time individually for metabolic labeling for…
Metabolic Labeling with AHA or HPG 下载为PDFGeneral Information For imaging, culture cells on cover slips or other desired imaging-compatible surface,
approximately 1 × 106 cells are sufficient for one detection. 1 × 106 cells are required to prepare
100 μg of lysate. Metabolic Labeling of Cells The following is a standa…
Labeling cells with EdU 下载为PDFThe following is a general protocol was tested with several cell lines including A549, HeLa, and NIH/3T3
cells, but it can be used for any adherent cell type. Growth medium, cell density, cell type variations, and
other factors may influence labeling. We recommend 10 μM E…
EdU Imaging, Flow Cytometry 下载为PDFThis protocol was tested with A549, HeLa, and NIH/3T3 cells but it can be adapted for any adherent cell
type. In initial experiments, we recommend starting with 1.5 µM final concentration of an azide detection
reagent. Slight adjustment of an azide detection reagent concentration might be r…
EdU Imaging, Microscopy 下载为PDFThis protocol was tested with A549, HeLa, and NIH/3T3 cells but it can be adapted for any adherent cell
type. In initial experiments, we recommend starting with 5 µM concentration of an azide detection
reagent. Slight adjustment of an azide detection reagent concentration might be r…
Cell Lysate Labeling 下载为PDF This is a general protocol for labeling cell lysate through copper-catalyzed click reaction with any azide or
alkyne click detection reagent. This protocol may be used as a starting point for optimization of particular
click c…
