The expression of CD44 and absence of CD24 (CD44+/CD24−) is characteristic of breast CSCs. To evaluate these, Hs578T and Hs578Ts(i)8 cell variants were seeded at 1 × 105 cells in a 6-well plate and allowed to attach overnight. They were subsequently trypsinised, blocked with 10% FCS in PBS and stained with APC-conjugated anti-CD24 (1:100) (eBioscience, San Diego, California, USA) and FITC-conjugated anti-CD44 (1:400) (eBioscience) for 30 min at 4 °C. Staining was assessed in a FACSCanto II flow cytometer, followed by analysis using BD FACSDiva software. To assess the effects of 2-DG on the CSC population Hs578T and Hs578Ts(i)8 cell variants were seeded at 1 × 105 cells in a 6-well plate and allowed to attach overnight. Cells were treated with 2-DG (final concentration 15 mM) for 24 hours. They were subsequently trypsinised, blocked with 10% FCS in PBS and stained with APC-conjugated anti-CD24 (1:100) (eBioscience, San Diego, California, USA) and FITC-conjugated anti-CD44 (1:400) (eBioscience) for 30 min at 4 °C. Staining was assessed in a FACSCanto II flow cytometer, followed by analysis using BD FACSDiva software.
Apc conjugated anti cd24
Manufactured by Thermo Fisher Scientific
Sourced in United States
APC-conjugated anti-CD24 is a fluorescently labeled monoclonal antibody that binds to the CD24 cell surface marker. It is used for the identification and analysis of cells expressing CD24 in flow cytometry applications.
Automatically generated - may contain errors
2 protocols using apc conjugated anti cd24
1
Evaluating Breast Cancer Stem Cells
2
CD24, CD44, and PD-L1 expression in breast cancer cell lines
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HCC1954 and SKBR3 cell variants (1 × 105/well) were seeded in a 6-well plate and allowed to attach overnight. They were then trypsinised, blocked with 10% FCS in PBS and stained with APC-conjugated anti-CD24 (1:75 dilution, eBiosciences, UK) and FITC-conjugated anti-CD44 (1:800 for HCC1954, 1:400 for SKBR3, eBiosciences, UK) or APC-conjugated anti-PD-L1 antibody (1:800 for HCC1954, 1:400 for SKBR3, eBiosciences, UK) for 30 min at 4°C. For EV treatment, cells were seeded at 0.5 × 105 (HCC1954) or 1 × 105 (SKBR3 and EFM192A) cells/well in a 24-well plate and allowed to attach overnight. The following day, cells were washed with serum-free media, fed with media supplemented with extracellular vesicle (EV)-depleted-FBS and treated with EVs for a further 48 hr before staining as above. Staining was assessed in a FACSCanto II flow cytometer, followed by analysis with BD FACSDiva software.
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