Primary pancreatic cells (monolayers and spheres) were trypsinized and resuspended in Sorting Buffer (3 μM EDTA, and 3% FBS in 1X PBS). To identify CD133 positive CSC, the following conjugated antibodies were used: anti-CD133/1-APC or PE; (Miltenyi), and appropriate isotype-matched control antibodies. For autofluorescent detection, cells were excited with blue laser 488 nm and selected as the intersection with the filters 530/30 (BL1) and 590/40 (BL2) [20 (link)]. For all assays, 2 mg/mL DAPI (Cat no. D9564, Sigma-Aldrich) was used to exclude dead cells with laser VL1. Data were analyzed with FlowJo 9.3 software (Tree Star Inc., Ashland, OR.). Cytometry data was acquired with an Invitrogen™ Attune™ NxT 4-laser cytometer with software version 3.1.1.
Dapi cat no d9564
Manufactured by Merck Group
DAPI (Cat no. D9564) is a fluorescent dye used for staining and visualizing nucleic acids, specifically DNA, in biological samples. It is a core laboratory tool commonly used in various applications such as fluorescence microscopy, flow cytometry, and high-content screening. The dye binds to DNA and emits a blue fluorescent signal when excited, allowing for the identification and localization of cell nuclei.
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Lab products found in correlation
2 protocols using dapi cat no d9564
1
Identification of Pancreatic Cancer Stem Cells
2
Isolation and Analysis of Pancreatic Cancer Stem Cells
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Primary pancreatic cells (monolayers and spheres) were trypsinized and resuspended in Sorting Buffer (3 µM EDTA, and 3 % FBS in 1X PBS). To identify CD133 positive CSC, the following conjugated antibodies were used: anti-CD133/1-APC or PE; (Miltenyi), and appropriate isotype-matched control antibodies. For auto uorescent detection, cells were excited with blue laser 488 nm and selected as intersection with the lters 530/30 (BL1) and 590/40 (BL2) (20) . For all assays, 2 mg/mL DAPI (Cat no. D9564, Sigma) was used to exclude dead cells with laser VL1. Data were analyzed with FlowJo 9.3 software (Tree Star Inc., Ashland, OR.). Cytometry data was acquired with an Invitrogen™ Attune™ NxT 4-laser cytometer with software version 3.1.1.
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