GBM cells were analyzed for common stem cell markers by flow cytometry. Briefly, cells were collected, washed twice with PBS 1x, and stained with APC-conjugated anti-CD44 antibody (dilution 1:10) (Becton Dickinson, BD Pharmigen, cat n. 559942 RRID: AB_398683) and PE-conjugated anti- CD24 antibody (dilution 1:20) (Becton Dickinson, BD Pharmigen, cat n. 555428, RRID: AB_395822) for 30 min at 4 °C. APC-conjugated anti- CD133 antibody (dilution 1:10) (Miltenyi Biotec, cat n. 130-090-826) was used for single staining. After three washes, fluorescence was acquired using a FACS Canto flow cytometer (Becton Dickinson, San Diego, CA, USA) equipped with 488 and 633 laser lines. Appropriate isotype control was included for each sample.
Apc conjugated anti cd133 antibody
Manufactured by Miltenyi Biotec
Sourced in Germany
The APC-conjugated anti-CD133 antibody is a flow cytometry reagent used for the identification and isolation of CD133-positive cells. CD133 is a cell surface antigen commonly used as a marker for stem and progenitor cells. The antibody is conjugated with Allophycocyanin (APC), a fluorescent dye, enabling the detection and quantification of CD133-expressing cells by flow cytometry.
Automatically generated - may contain errors
Lab products found in correlation
Facscanto flow cytometer, by BD (1 mentions)
Pe conjugated anti cd11b, by BD (1 mentions)
Apc conjugated anti cd235a, by BD (1 mentions)
Rhil 9, by Thermo Fisher Scientific (1 mentions)
Anti cd43, by BD (1 mentions)
Rhil 11, by Thermo Fisher Scientific (1 mentions)
Anti filamin a, by Santa Cruz Biotechnology (1 mentions)
Accucount fluorescent particles, by Spherotech (1 mentions)
Anti cd42b, by BD (1 mentions)
7 aad labeling, by BD (1 mentions)
Vioblue conjugated anti cd34 antibody, by Miltenyi Biotec (1 mentions)
Anti cd3 monoclonal antibody, by Miltenyi Biotec (1 mentions)
Accutase, by STEMCELL (1 mentions)
4 protocols using apc conjugated anti cd133 antibody
1
Stem Cell Marker Analysis in GBM Cells
2
Characterization of Platelet-Derived Extracellular Vesicles
3
Phenotypic and Transduction Efficiency of Stem Cells and T Cells
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The phenotype and efficiency of transduction of hCD34+ HSPCs and T lymphocytes were analyzed by Flow Cytometry (FACS Canto II Instrument and DIVA software). hCD34+ HSPCs were stained with PE-conjugated anti-CD90 antibody (BD PharMingen), VioBlue-conjugated anti-CD34 antibody (Miltenyi Biotec), and antigen-presenting cell (APC)-conjugated anti-CD133 antibody (Miltenyi Biotec).
PBMCs were stained with anti-CD3 monoclonal antibody before and after activation (Miltenyi Biotec). In selected experiments, T cell memory phenotype was detected by staining with monoclonal antibodies specific for CD3 (VioGreen-conjugated), CD8 (APC-Vio770-conjugated), CD45RA (PE-Vio770-conjugated), CD62L (VioBlue-conjugated), and CD95 (APC-conjugated) (Miltenyi Biotec). For both cell types, cell mortality and transduction efficiency were evaluated by 7-AAD labeling (BD PharMingen) and by the level of GFP expression, respectively.
PBMCs were stained with anti-CD3 monoclonal antibody before and after activation (Miltenyi Biotec). In selected experiments, T cell memory phenotype was detected by staining with monoclonal antibodies specific for CD3 (VioGreen-conjugated), CD8 (APC-Vio770-conjugated), CD45RA (PE-Vio770-conjugated), CD62L (VioBlue-conjugated), and CD95 (APC-conjugated) (Miltenyi Biotec). For both cell types, cell mortality and transduction efficiency were evaluated by 7-AAD labeling (BD PharMingen) and by the level of GFP expression, respectively.
4
Quantifying GBM Cancer Stem Cells
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The subpopulation of cancer stem cells in GBM tumor spheroids or cultured adherently were sorted and evaluated with and without enzalutamide treatment, respectively, with an anti-CD133 CSC surface antibody (Miltenyi Biotec, Germany). After treating the tumor spheroids for 4 days with 120 µM enzalutamide or 180 µM bicalutamide, the tumor spheroids were harvested, gently dissociated to single cell suspensions using ACCUTASE™ (STEMCELL Technologies Inc., Canada). First a total of 106 cells were stained with Live/Dead fixable dead cell staining dyes and then incubated with APC-conjugated anti-CD133 antibody (Miltenyi Biotec, Germany). After 30 min of incubation with CD133 antibody at 4°C, cells were washed with phosphate-buffered saline (PBS) at 300×g for 10 min. Samples were sorted using a FACS LSRII G Flow Cytometer and percentages of CSC subpopulation were analyzed by FACSDiva software (Beckon-Dickinson, Franklin Lakes, NJ, USA).
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