Cells were acquired using the Cytofix/CytopermTM Fixation/Permeabilization Solution Kit (BD Biosciences), and then the percentage of Ki67-positive cells was determined. Anti-Ki67 antibody was added (1:500), and the group of tubes without antibody was used as the control. Antibody against Ki67 and antibody against rabbit immunoglobin G (IgG); H+L; Phycoerythrin (PE) were obtained from Abcam.
Phycoerythrin pe
Manufactured by Abcam
Phycoerythrin (PE) is a fluorescent protein derived from red algae. It serves as a highly efficient fluorescent label, with the ability to emit bright, stable fluorescence when excited by a suitable light source. PE has a wide range of applications in various experimental techniques, such as flow cytometry, immunoassays, and fluorescence microscopy.
Automatically generated - may contain errors
Lab products found in correlation
Cytofix cytoperm fixation permeabilization solution kit, by BD (1 mentions)
Lsm 5 pascal, by Zeiss (1 mentions)
Fluorescein isothiocyanate (fitc), by Abcam (1 mentions)
Oct compound, by Thermo Fisher Scientific (1 mentions)
Antifade solution, by Merck Group (1 mentions)
β catenin, by Abcam (1 mentions)
Foxj1, by Abcam (1 mentions)
2 protocols using phycoerythrin pe
1
Quantifying Cell Proliferation by Ki67
2
Immunofluorescent Characterization of Organoid Cultures
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Organoids were embedded in an optimum cutting temperature (OCT) compound (Thermo
Fisher Scientific, MA, USA) and stored at −80oC. The OCT-embedded
organoids were sectioned at 5 μm thickness. The sections were removed from the
OCT, washed with phosphate-buffered saline (PBS), and fixed with 4%
paraformaldehyde for 15 min. The sections were permeabilized with 0.1% Triton
X-100, blocked with 5% FBS, and treated with primary antibodies (1:100, FOXJ1
[ciliated cell marker], 1:100, PAX8 [secretory cell marker], and 1:100,
beta-catenin, all purchased from Abcam) for 1.5 h. The sections were washed with
PBS and 0.1% Tween 20, then treated with secondary antibodies [Fluorescein
isothiocyanate (FITC) or phycoerythrin (PE); Abcam] for 1 h. The sections were
then mounted in antifade solution (Sigma-Aldrich) and washed with PBS plus 0.1%
Tween 20. The nuclei were counterstained with DAPI, and the slides were
subjected to confocal laser-scanning microscopy (LSM5 PASCAL; Carl Zeiss, Jena,
Germany).
Fisher Scientific, MA, USA) and stored at −80oC. The OCT-embedded
organoids were sectioned at 5 μm thickness. The sections were removed from the
OCT, washed with phosphate-buffered saline (PBS), and fixed with 4%
paraformaldehyde for 15 min. The sections were permeabilized with 0.1% Triton
X-100, blocked with 5% FBS, and treated with primary antibodies (1:100, FOXJ1
[ciliated cell marker], 1:100, PAX8 [secretory cell marker], and 1:100,
beta-catenin, all purchased from Abcam) for 1.5 h. The sections were washed with
PBS and 0.1% Tween 20, then treated with secondary antibodies [Fluorescein
isothiocyanate (FITC) or phycoerythrin (PE); Abcam] for 1 h. The sections were
then mounted in antifade solution (Sigma-Aldrich) and washed with PBS plus 0.1%
Tween 20. The nuclei were counterstained with DAPI, and the slides were
subjected to confocal laser-scanning microscopy (LSM5 PASCAL; Carl Zeiss, Jena,
Germany).
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