Selected article for: "calcium phosphate and precipitation method"
Title: Localization of TGN38 to the trans-Golgi network: involvement of a cytoplasmic tyrosine-containing sequence
  • Document date: 1993_3_1
    • ID: qt44izzh_15
    Snippet: Cells grown to 40-50% confluence on glass coverslips were transiently transfected with 5 /~g of DNA by the calcium phosphate precipitation method, as described above. After 36 to 60 h, cells were fixed for 15 rain at room temperature in 2% formaldehyde in PBS and washed in PBS. The cells were then incubated for 1 h with primary antibodies in PBS containing 0.1% BSA and 0.2% saponin, washed in PBS, incubated with fluorescently labeled secondary an.....
    Document: Cells grown to 40-50% confluence on glass coverslips were transiently transfected with 5 /~g of DNA by the calcium phosphate precipitation method, as described above. After 36 to 60 h, cells were fixed for 15 rain at room temperature in 2% formaldehyde in PBS and washed in PBS. The cells were then incubated for 1 h with primary antibodies in PBS containing 0.1% BSA and 0.2% saponin, washed in PBS, incubated with fluorescently labeled secondary antibodies for 30 min, washed again in PBS, and then mounted onto glass slides with Fluoromount G (Southern Biotechnology Associates, Birmingham, AL). Samples were examined under a Zeiss inverted microscope equipped with a 63 x lens (Carl Zeiss, Oberkochen, Germany). The same immunofluorescent labeling procedure was used for staining stably transfected cells.

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