Total tumor cells for each individual sample were subjected to magnetic cell sorting independently using CD44 (anti-human) MicroBeads (Miltenyi Biotec, Bergisch Gladbach, Germany) per the protocol provided by the manufacturer. The sorted CD44+ cells were stained with Anti-CD44-PE antibody (Miltenyi Biotec, Auburn, CA, USA) and acquired on a flow cytometer (Attune NxT, Thermo Fisher Scientific, Waltham, MA, USA) to check the purity and seeded with complete media (DMEM + 10% FBS) for further experimentation.
Anti cd44 pe antibody
Manufactured by Miltenyi Biotec
Sourced in United States
The Anti-CD44-PE antibody is a fluorescently labeled antibody that specifically recognizes the CD44 cell surface receptor. CD44 is involved in various cellular processes, including cell-cell interactions, cell adhesion, and migration. The PE (Phycoerythrin) fluorescent label allows for the detection and analysis of CD44-expressing cells using flow cytometry or other fluorescence-based techniques.
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2 protocols using anti cd44 pe antibody
1
CD44+ Cell Isolation and Validation
2
ALDEFLUOR Assay for Stem Cell Identification
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The organoids were collected by centrifugation at 300g for 5min, and dissociated with trypsin-EDTA for 5min at 37°C. The cells were incubated with ALDEFLUOR staining (STEMCELL TECHNOLOGIES) for 30 minutes at 37°C, and then anti-CD44-PE antibody (1:50 Miltenyi) for 25 minutes on ice in ALDEFLUOR Assay Buffer according to the protocol. The samples with DEAB inhibitor without CD44-PE staining serve as the negative control for ALDEFLUOR. For chemo dose response evaluation and CSL inhibitor screening, each sample takes 200 µL /mL for ALDEFLUOR Assay and 100 µL ALDEFLUOR Assay Buffer for CD44 staining. DAPI (1 µg/mL for final concentration) was added to label the dead cells before FACS test.
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