PMC:1914094 / 35894-36737 JSONTXT

{"target":"https://pubannotation.org/docs/sourcedb/PMC/sourceid/1914094","sourcedb":"PMC","sourceid":"1914094","source_url":"http://www.ncbi.nlm.nih.gov/pmc/1914094","text":"Immunofluorescence microscopy\nHeLa cells were seeded on coverslips and transfected with expression constructs using CaCl2 precipitation. After 2 days, transfected cells were fixed with 3% formaldehyde and 2% sucrose in PBS for 7 min at RT. Fixed cells were washed three times with PBS and incubated with different Trx1 constructs or recombinant CD30L (R\u0026D Systems). Proteins were visualized using appropriate primary antibodies (Anti-CD30L polyclonal antibody (R\u0026D Systems), anti-CD30 monoclonal antibodies Ki-1 (Santa Cruz) or Ber-H2 (DakoCytomation), anti-CD95 monoclonal antibody (a kind gift from Dr P Krammer), anti-Trx1 polyclonal antibody (M Preuss and TP Dick, unpublished) followed by FITC-conjugated anti-goat IgG, FITC-conjugated anti-rabbit IgG or TRITC-conjugated anti-mouse IgG and analyzed with a Nikon C1Si confocal microscope.","divisions":[{"label":"Title","span":{"begin":0,"end":29}}],"tracks":[]}