Selected article for: "confocal microscope and primary antibody"
Author: Claus, Claudia; Manssen, Lena; Hübner, Denise; Roßmark, Sarah; Bothe, Viktoria; Petzold, Alice; Große, Claudia; Reins, Mareen; Mankertz, Annette; Frey, Teryl K.; Liebert, Uwe G.
Title: Activation of the Mitochondrial Apoptotic Signaling Platform during Rubella Virus Infection
  • Document date: 2015_11_26
    • ID: 1rrm4sqa_59
    Snippet: Cells were fixed with paraformaldehyde (2% w/v), permeabilized with methanol, 0.1% Triton X-100, 0.3% Tween 20, or digitonin as indicated. Unspecific binding was blocked through incubation with 5% goat serum for 1 h at 37 ¥ C. After intensive washing incubation with primary antibody (the anti-E1 antibody was used at a 1:200 dilution in PBS, anti-CytC at 1:250, anti-CypA and anti-alpha tubulin at 1:500, all other antibodies at 1:100) was performe.....
    Document: Cells were fixed with paraformaldehyde (2% w/v), permeabilized with methanol, 0.1% Triton X-100, 0.3% Tween 20, or digitonin as indicated. Unspecific binding was blocked through incubation with 5% goat serum for 1 h at 37 ¥ C. After intensive washing incubation with primary antibody (the anti-E1 antibody was used at a 1:200 dilution in PBS, anti-CytC at 1:250, anti-CypA and anti-alpha tubulin at 1:500, all other antibodies at 1:100) was performed for 1 h at 37 ¥ C. Secondary antibodies (DyLight 488-or Cy3-labelled, Dianova GmbH (Hamburg, Germany) were used at a 1:100 dilution and incubated at 37 ¥ C for 45 min, thereafter as nuclear counterstain Hoechst bisbenzamide 33285 (5 µg/mL, Thermo Fisher Scientific) was applied. As a mitochondrial counterstain, MitoTracker Red CMXRos was applied according to the instructions of the MitoProbe transition pore opening kit. Samples were mounted in Entellan and images were taken with the Zeiss510 confocal microscope (Zeiss, Oberkochen, Germany) and processed using CorelDRAW X7 (Corel Corporation, Ottawa, Ontario, Canada) with minimal alterations to brightness and contrast.

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