MCF-7 CSCs were obtained by sorting as reported previously (Fu et al., 2017 (link)). In brief, the cells were concentrated to a few million/100 μL, and were stained by adding 10 μL anti-CD44-PE antibody (BD Pharmingen) and 10 μL anti-CD24-FITC antibody (BD Pharmingen). After the staining was completed, the cells were washed twice by PBS and analyzed/sorted on flow cytometer (BD Aria III).
Anti cd44 pe antibody
Manufactured by BD
The Anti-CD44-PE antibody is a fluorescently labeled antibody that binds to the CD44 cell surface receptor. CD44 is a common marker expressed on various cell types. This antibody can be used for the identification and analysis of CD44-positive cells in flow cytometry applications.
Automatically generated - may contain errors
Lab products found in correlation
Anti cd24 fitc antibody, by BD (1 mentions)
Aria 3, by BD (1 mentions)
Facsverse cytometer, by BD (1 mentions)
Propidium iodide, by Merck Group (1 mentions)
Cellquest software program, by BD (1 mentions)
Facscalibur flow cytometer, by BD (1 mentions)
Anti cd24 apc, by BD (1 mentions)
Facsaria cell sorter, by BD (1 mentions)
5 protocols using anti cd44 pe antibody
1
Isolation of MCF-7 Cancer Stem Cells
2
Breast Cancer Stem Cell CD44 Profiling
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To examine the breast cancer stem cell population with enriched CD44 expression, the LDR-exposed 5 × 105 breast cancer cells were labeled with an anti-CD44/PE antibody (BD Biosciences, Korea). A respective control was also prepared and tested for each sample. Briefly, control and exposed cells were incubated for 20–25 min at 4 °C, washed twice with PBS, and immediately analyzed using a BD FACSVerse cytometer and the FACS suite software. The percentage of dead cells was determined using propidium iodide (PI, 50 ng/ml) (Sigma) according to the manufacturer’s instructions and was analyzed using a flow cytometer.
3
Flow Cytometric Analysis of CD44 Expression
4
ACK1 Inhibitors and Prostate Cancer
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VCaP and LNCAP cell lines were obtained from ATCC. LNCaP-CaAck cells were developed, as described earlier36 (link). ACK1 monoclonal Ab (A11), actin, phosphotyrosine and AR monoclonal antibodies were purchased from Santacruz; Anti-phospho-ACK1 (Tyr284, Upstate) were purchased from Cell Signaling. Anti-CD44-PE antibodies were purchased from BD Biosciences. (R)-9b MS and AIM-100 were synthesized at Moffitt Cancer Center as described earlier27 (link). Control and ACK1 siRNAs were generated by custom synthesis (Qiagen) and the sequences have been described previously20 (link). For immunoprecipitations, cells were lysed in receptor lysis buffer (RLB) containing 25 mmol/L Tris (pH 7.5), 500 mmol/L NaCl, 1% Triton X-100, 10% glycerol, phosphatase inhibitors (10 mmol/L NaF, 1 mmol/L Na2VO4), and protease inhibitor mix (Roche).
5
Identification of Cancer Stem Cell Markers
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Anti-CD24-APC and anti-CD44-PE antibodies were purchased from BD Biosciences. Anti-CD133-Biotin and anti-Alpha-2 Beta-1 (α2β1) Integrin-FITC antibody were purchased from Miltenyi Biotech and Abcam, respectively. Cell labeling with fluorescent-conjugated antibodies was performed according to the manufacturer's recommendations. Sorting of antibody-labeled cells was carried out on a FACSAria cell sorter (BD Biosciences). Sorted CD24 -CD44 + cells from LNCaP or DU145cells were cultured in MEM containing 10% fetal bovine serum and insulin (10 μg/ml) at 37°C with 5% CO 2 . Sorted CD24 -CD44 + and CD24 + CD44 - cells were subsequently tested for expression of CD133 and α2β1 Integrin.
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