Super-resolution Airyscan images were acquired on a Zeiss LSM 980 with Airyscan microscope (Carl Zeiss). Samples were prepared on the CellCarrier 96 ultral microplate (PerkinElmer). TOMM20 and dsDNA were labelled with monoclonal rabbit antibodies against TOMM20 (Abcam, ab232589, 1:500) and monoclonal mouse antibodies against dsDNA (Sigma-Aldrich, MAB1293, 1:500), respectively. The samples were then washed and incubated with Alexa Fluor 568/488 secondary antibodies. The nucleus was stained with Hoechst 33342 (Sigma-Aldrich, B2261). Data were collected using a ×63/1.4 NA objective for the majority of experiments, under the control of the Zeiss ZEN software; 405, 488 and 561 nm laser lines were used. In cases in which z-stacks were collected, the software-recommended optimal slice sizes were used. Airyscan image processing was performed using the Airyscan processing function with the standard mode in the ZEN software. To maintain clarity and uniformity throughout the paper, the images were pseudocoloured.
Ab232589
Manufactured by Abcam
Ab232589 is a laboratory equipment product offered by Abcam. It serves as a general-purpose tool for researchers in various scientific fields. The core function of this product is to provide a standardized and reliable mechanism for conducting experimental procedures.
Automatically generated - may contain errors
Lab products found in correlation
Lsm 980, by Zeiss (1 mentions)
Hoechst 33342, by Merck Group (1 mentions)
Airyscan microscope, by Zeiss (1 mentions)
Mab1293, by Merck Group (1 mentions)
Donkey serum, by Jackson ImmunoResearch (1 mentions)
Tunel assay kit, by Abcam (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (1 mentions)
Paraformaldehyde (pfa), by Santa Cruz Biotechnology (1 mentions)
Magnesium chloride, by Thermo Fisher Scientific (1 mentions)
Dulbecco s pbs, by Thermo Fisher Scientific (1 mentions)
2 protocols using ab232589
1
Super-resolution Imaging of Mitochondria and DNA
2
Immunofluorescence Staining of 2D Cells
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Cells grown in 2D were rinsed with Dulbecco's PBS with calcium chloride and magnesium chloride (Gibco) before being fixed for 20 min in a cold 4% paraformaldehyde (ChemCruz) solution. The cells were blocked and permeabilized in 10% Donkey Serum (Jackson ImmunoResearch)/0.1% Triton-X (Sigma) for 30 min at room temperature. The cells were then incubated overnight at 4 °C with the primary antibody (diluted 1:100 in 1% Donkey Serum/0.1% Triton-X) [53 ]. Subsequently, the cells were incubated with the secondary antibody (diluted 1:1000 in 1% Donkey Serum/0.1% Triton-X) for 1 h at room temperature. For nuclear staining, cells were incubated with 4′,6-diamidino-2-phenylindole (DAPI, 1:5000 in Dulbecco's PBS; Thermo Scientific) for 5 min at room temperature. A primary antibody for Tom 20 (#ab232589, Abcam) was used at a 1:500 dilution. TUNEL staining was performed with a TUNEL Assay Kit (FITC; #ab66108, Abcam) as we previously described [14 ].
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