Flow cytometry analysis was performed using anti-SSEA1-eFluor 660 (eBioscience 50-8813-42, 1:20), anti-CD44-FITC (IM7) (eBioscience, 11-0441-81, 0.5 mg per test) and anti-CD24-VioBlue (clone: 32D12) (Miltenyi biotech, 130-099-150, 1:11). For tumor-derived samples, anti-mouse MHC Class I H2-Kd (eBioscience 17-5957-82, 1:200) followed by PerCP-Cy5.5-conjugated streptavidin (eBioscience 45-4317-80, 1:200) was used to exclude host cells. Cells were incubated with antibodies at a concentration of 1 × 106 cells/100 μL in sort buffer (15 mM HEPES buffer [Sigma H3375], 1% Bovine Serum Albumin [Sigma A9647], 2 mM EDTA [VWR 20302.260], 100 U/mL DNase I (NEB M0303), 100 U/mL penicillin and 100 μg/mL streptomycin [Thermo Fisher 15140122] in PBS) for 45 min on ice, washed 3 times with sort buffer and analyzed using LSR Fortessa (BD Biosciences) or sorted using FACSAria II (BD Biosciences). FACSDiva and FlowJo software were used to acquire and analyzed data, respectively. Gating strategy used for tumor analysis is shown in Supplementary Fig. 1 .
Anti cd24 vioblue clone 32d12
Manufactured by Miltenyi Biotec
Anti-CD24-VioBlue (clone: 32D12) is a monoclonal antibody that binds to the CD24 antigen. CD24 is a glycosylphosphatidylinositol (GPI)-anchored cell surface protein expressed on a variety of cell types. The anti-CD24-VioBlue antibody is conjugated with the VioBlue fluorochrome, which can be used for flow cytometric analysis.
Automatically generated - may contain errors
Lab products found in correlation
Penicillin, by Thermo Fisher Scientific (4 mentions)
Streptomycin, by Thermo Fisher Scientific (4 mentions)
Dnase 1, by New England Biolabs (2 mentions)
Anti ssea1 efluor 660, by Thermo Fisher Scientific (2 mentions)
Anti cd44 fitc im7, by Thermo Fisher Scientific (2 mentions)
Percp cy5.5 conjugated streptavidin, by Thermo Fisher Scientific (2 mentions)
Facsdiva, by FlowJo (2 mentions)
Bovine serum albumin (bsa), by Merck Group (2 mentions)
Lsrfortessa, by BD (2 mentions)
Facsaria 2, by BD (2 mentions)
2 protocols using anti cd24 vioblue clone 32d12
1
Flow Cytometry Analysis of SSEA1, CD44, and CD24
2
Flow Cytometry Analysis of SSEA1, CD44, and CD24
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Flow cytometry analysis was performed using anti-SSEA1-eFluor 660 (eBioscience 50-8813-42, 1:20), anti-CD44-FITC (IM7) (eBioscience, 11-0441-81, 0.5 mg per test) and anti-CD24-VioBlue (clone: 32D12) (Miltenyi biotech, 130-099-150, 1:11). For tumor-derived samples, anti-mouse MHC Class I H2-Kd (eBioscience 17-5957-82, 1:200) followed by PerCP-Cy5.5-conjugated streptavidin (eBioscience 45-4317-80, 1:200) was used to exclude host cells. Cells were incubated with antibodies at a concentration of 1 × 106 cells/100 μL in sort buffer (15 mM HEPES buffer [Sigma H3375], 1% Bovine Serum Albumin [Sigma A9647], 2 mM EDTA [VWR 20302.260], 100 U/mL DNase I (NEB M0303), 100 U/mL penicillin and 100 μg/mL streptomycin [Thermo Fisher 15140122] in PBS) for 45 min on ice, washed 3 times with sort buffer and analyzed using LSR Fortessa (BD Biosciences) or sorted using FACSAria II (BD Biosciences). FACSDiva and FlowJo software were used to acquire and analyzed data, respectively. Gating strategy used for tumor analysis is shown in Supplementary Fig. 1 .
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