At P3, cells were harvested by trypsinization with 0.05% trypsin (Invitrogen) upon reaching 90% confluence, and re-suspended in DPBS to reach a final cell density of 1.5 × 106 cells/ml. An amount of 200 μl of cell suspension (1 × 105 cells) was incubated in the dark for 1 hr at 37°C with Phycoerythrin-conjugated antibodies against CD44, CD73, CD166, CD105 and CD34, and fluoro isothycyanate-conjugated antibodies against CD45 and HLA-DR (all from BD Pharmingen) for specific surface antigens analysis by using flow cytometer. Excess antibodies were removed by washing with DPBS. All analyses were standardized against negative control cells incubated with Isotype-specific IgG1-PE and IgG1-FITC (BD Pharmingen). At least 10,000 events were acquired on Guava Technologies flow cytometer, and the results were analysed by using Cytosoft, Version 5.2, Guava Technologies.
Phycoerythrin conjugated antibodies against cd44
Manufactured by BD
Sourced in United States
Phycoerythrin-conjugated antibodies against CD44 are a type of fluorescent-labeled antibody used in flow cytometry and other immunoassays. These antibodies bind specifically to the CD44 cell surface receptor, which is involved in cell-cell interactions, cell adhesion, and cell migration. The phycoerythrin dye conjugated to the antibody emits a bright orange-red fluorescence when excited by a laser, allowing for the detection and quantification of CD44-expressing cells.
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2 protocols using phycoerythrin conjugated antibodies against cd44
1
Mesenchymal Stem Cell Surface Marker Profiling
2
Intravenous Transplantation of hAFSCs in Diabetic Rats
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hAFSCs were obtained by routine amniocentesis from six healthy pregnant donors in 15-20 gestational weeks. Cells were cultured in the StemPro mesenchymal stem cell (MSC) serum free medium supplemented with 10% fetal bovine serum (Invitrogen, Carlsbad, CA, United States) and incubated at 37 °C with 5% carbon dioxide. The specific surface antigens of hAFSCs were characterized by flow cytometry analyses as shown in previous work[18 (link)]. The cultured cells were trypsinized and stained with phycoerythrin-conjugated antibodies against CD44, CD73, CD90, CD105, CD117, and CD45 (BD PharMingen, CA, United States). Thereafter, the cells were analyzed using the Calibur flow cytometer (Becton Dickinson, Heidelberg, Germany). hAFSCs at passages 4-6 were collected and prepared to a final concentration of 3 × 106 cells/mL phosphate buffered saline. In the DM + hAFSCs group, a single dose of 3 × 106 hAFSCs was injected intravenously via the tail vein at 7 d after DM induction. The treatment dose of hAFSCs was determined according to a previous study that used adipose tissue-derived stem cells to treat bladder dysfunction of STZ-induced diabetic rats[10 (link)].
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