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Pe conjugated cd44

Manufactured by BioLegend
Sourced in United States

PE-conjugated CD44 is a fluorescently-labeled antibody that binds to the CD44 cell surface antigen. CD44 is a transmembrane glycoprotein involved in cell-cell interactions, cell adhesion and migration. The PE (phycoerythrin) fluorescent dye is conjugated to the CD44 antibody, allowing for the detection and analysis of CD44-expressing cells using flow cytometry or other fluorescence-based techniques.

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2 protocols using pe conjugated cd44

1

Comprehensive Erythroid Cell Analysis

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A modified version of an erythroid differentiation protocol was used for flow cytometric analysis (Liu et al, 2013). Antibodies used were phycoerythrin‐cyanin 7 (PECy7)‐conjugated TER119 (eBioscience, San Diego, CA, USA), and phycoerythrin (PE)‐conjugated CD44 (BioLegend, San Diego, CA, USA). Allophycocyanin (APC)‐conjugated B220, CD3 and GR‐1 (BioLegend) were used as lineage markers. 7‐aminoactinomycin D (7AAD) was used to monitor viability. The plots were gated on live cells of appropriate size, negative for lineage markers. For flow cytometric detection of apoptosis, PE‐conjugated AnnexinV apoptosis detection kit (BD biosciences, Franklin Lakes, NJ, USA) was used according to the manufacturer's instructions. Data collection was done on a Canto II flow cytometer (BD Biosciences). For cell cycle analysis, the cells were disrupted in ice‐cold ethanol and the DNA was stained with propidium iodide. Cell cycle analysis was done on Aria III flow cytometer (BD Biosciences). All analyses were performed using FlowJo v10 (Tree Star Inc., San Carlos, CA, USA).
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2

Characterization of Canine Cell Phenotypes

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The surface markers of isolated cells were analyzed by flow cytometry analysis. Briefly, the isolated cells (1 × 106 cells, passage 3) from the three dogs were, respectively, suspended in 100 μL phosphate-buffered saline (PBS) containing 10 μg/mL antibodies for PE-conjugated CD29 (303003, BioLegend, USA), PE-conjugated CD44 (103024, BioLegend, USA), PE-conjugated CD90 (561970, BD Biosciences, USA), PE-conjugated CD105 (bs-0579R-PE, Bioss, CHN), FITC-conjugated CD73 (bs-23233R-FITC, Bioss, CHN), PE-conjugated CD34 (559369, BD Biosciences, USA), and FITC-conjugated CD45 (11-5450-42, eBioscience, USA). After incubation for 30 min at 4°C, the cells were washed with PBS and then resuspended in 500 μL of PBS for analysis. As for CD11b, the isolated cells (1 × 106 cells, passage 3) were suspended in 100 μL phosphate-buffered saline (PBS) containing 10 μg/mL CD11b (MA5-16604, eBioscience, USA). After incubation for 30 min at 4°C, the cells were washed with PBS and then labeled with goat antirabbit IgG (H + L) cross-adsorbed secondary antibody, FITC (F-2765, Invitrogen), at a dilution of 1 : 500 for 1 h at room temperature. Cell fluorescence was evaluated by flow cytometry using a DxP Athena™ flow cytometry system (Cytek) and analyzed with FlowJo 10 software (Tree Star, USA).
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