Staining for Ki67 and phospho-histone H3 (H3P) was performed on trypsinized cells from NC and CD organoids previously fixed in formaldehyde 4% and permeabilized in methanol 90%. Cells were then washed in PBS, incubated with anti-Ki67 antibody (FITC-conjugate, #16667, Abcam) and H3P (Pacific Blue conjugate, #8552, Cell Signaling Technology, USA) in Bovine Serum Albumin 0.5% in PBS (#A7906, Sigma-Aldrich) and washed twice. 105 cells per samples were acquired and analysis was performed on gated cells that excluded debris and double positive (Ki67+H3P+) representative of cells that entered mitosis. Data were acquired with BD FACS Calibur flow cytometer instrument. Analysis was performed with BD CellQuestPro software.
Fitc conjugate
Manufactured by Abcam
Sourced in United States, United Kingdom
FITC-conjugate is a fluorescent label used in various biomedical applications. It is a commonly used fluorescent dye that emits green fluorescence when excited by a suitable light source. The core function of FITC-conjugate is to serve as a labeling agent for the detection and visualization of biomolecules, such as proteins, antibodies, or nucleic acids, in a wide range of analytical techniques.
Automatically generated - may contain errors
2 protocols using fitc conjugate
1
Quantifying Mitotic Cell Fraction
2
Macrophage Phenotyping by Flow Cytometry
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Markers for M1-type (CD86) macrophages and M2-type (CD206) macrophages were assessed by flow cytometry. Macrophages were incubated with anti-CD86 (20 μL/106 cells, FITC Conjugate; Abcam, Cambridge, UK) and anti-CD206 (10 μL/106 cells, PE-A Conjugate; Proteintech) for 30 min at 4°C in the dark. After that, CD86 positive cells and CD206 positive cells were detected using a CytoFLEX flow cytometer (Beckman Coulter, Indianapolis, IN, USA).
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