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Cd133 2 pe

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The CD133/2-PE is a fluorescence-labeled antibody that binds to the CD133 antigen. CD133 is a pentaspan transmembrane glycoprotein that is expressed on various stem and progenitor cells. The PE (phycoerythrin) fluorochrome is used to label the antibody, allowing for the detection and analysis of CD133-positive cells using flow cytometry or other fluorescence-based techniques.

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Lab products found in correlation

Facscalibur machine, by BD (1 mentions) Facs lysing solution, by BD (1 mentions) Facsaria flow cytometer, by BD (1 mentions) Facsdiva, by BD (1 mentions) Stempro accutase cell dissociation reagent, by Thermo Fisher Scientific (1 mentions) Permeabilization solution 2, by BD (1 mentions) Lsrii flow cytometer, by BD (1 mentions) Facsdiva software, by BD (1 mentions) Aldefluor assay, by STEMCELL (1 mentions) Annexin 5, by Thermo Fisher Scientific (1 mentions) Mirvana mirna isolation kit, by Thermo Fisher Scientific (1 mentions) Collagenase enzyme, by Thermo Fisher Scientific (1 mentions) Fixation permeabilization solution kit, by BD (1 mentions) Cd166 pe, by Thermo Fisher Scientific (1 mentions) Cxcr4 pe, by Thermo Fisher Scientific (1 mentions) Cd9 fitc, by Thermo Fisher Scientific (1 mentions) Dxp10, by Cytek (1 mentions) Annexin 5 fitc apoptosis detection kit, by Thermo Fisher Scientific (1 mentions) Cd44 pecy5, by Thermo Fisher Scientific (1 mentions) Cd15 apc, by BD (1 mentions)

Spelling variants of this product

CD133 (122 citations) CD133-PE (52 citations) CD133/2 (293C3)-PE antibody (5 citations) CD133/2-PE antibody (5 citations) Anti-CD133/2-PE (4 citations) CD133/2 (293C3)-PE (3 citations) Mouse anti-human CD133/2-PE (2 citations) PE-conjugated antihuman CD133/2 monoclonal antibody (mAb) (2 citations) Anti-CD133/2-PE antibody (2 citations)

8 protocols using cd133 2 pe

1

Isolation of CD133+ Cancer Cells

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Cells were first irradiated with 4 Gy and then treated with 10 μM RO4929097 and 10 μM TMZ as well as their combination. Cultures were disaggregated with accutase 4 days post-treatment, cells were washed with PBS once and incubated with CD133/2-PE (1:200; Miltenyi Biotech) or isotype control antibody on ice for 30 min. Then cells were washed with PBS 2×, resuspended in 200 μl PBS and fixed in 1% paraformaldehyde. Just before measurements of each FACS tube DAPI (final concentration 3 μM) was resuspended in each tube. FACS analysis was carried out on FACS Calibur machine (BD Biosciences).
Yahyanejad S., King H., Iglesias V.S., Granton P.V., Barbeau L.M., van Hoof S.J., Groot A.J., Habets R., Prickaerts J., Chalmers A.J., Eekers D.B., Theys J., Short S.C., Verhaegen F, & Vooijs M. (2016). NOTCH blockade combined with radiation therapy and temozolomide prolongs survival of orthotopic glioblastoma. Oncotarget, 7(27), 41251-41264.
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2

CD133 Expression Profiling of Stem Cells

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Subspheres were harvested with StemProAccutase Cell Dissociation Reagent (Thermo Fisher Scientific, Carlsbad, CA) and washed with PBS (pH = 7.4). For intracellular staining, the cells were fixed (FACS Lysing Solution, BD Biosciences) and permeabilized (Permeabilization Solution 2, BD Biosciences, San Jose, CA). Human monoclonal antibody CD133/2 PE (clone: 133/2; Miltenyi Biotec, Bergisch Gladbach, Germany) (BD Biosciences, San Diego, CA) was used. The data were acquired with a FACSAria flow cytometer (BD Biosciences, San Jose, CA) and analyzed by using FACSDiva (BD Biosciences, San Jose, CA) or FlowJo software (Tree Star, Ashland, OR).
Pavon L.F., Sibov T.T., de Souza A.V., da Cruz E.F., Malheiros S.M., Cabral F.R., de Souza J.G., Boufleur P., de Oliveira D.M., de Toledo S.R., Marti L.C., Malheiros J.M., Paiva F.F., Tannús A., de Oliveira S.M., Chudzinski-Tavassi A.M., de Paiva Neto M.A, & Cavalheiro S. (2018). Tropism of mesenchymal stem cell toward CD133+ stem cell of glioblastoma in vitro and promote tumor proliferation in vivo. Stem Cell Research & Therapy, 9, 310.
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3

Quantifying Stemness and Apoptosis

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The ALDH activity in live cells was measured by an ALDEFLUOR Assay (Stem Cell Technologies, Catalog No. 01700, Vancouver, BC, Canada) as per the manufacturer’s protocol. The percentage of ALDH+ cells was determined by an LSRII flow cytometer (BD Biosciences, Franklin Lakes, NJ, USA), using 488 nm excitation, and the signal was detected using the 530/30 filter. The ALDH+ percentage gate was determined by a sample-specific negative control diethylamino benzaldehyde (DEAB)/ALDH+ gate. CD133 was detected by flow cytometry using the fluorescent-labelled antibody CD133/2-PE (Miltenyi Biotec, Catalog No. 130-120-145, San Diego, CA, USA) in an LSRII flow cytometer using the filter 582/15. For each experiment, 10,000 events were analyzed. PI/Annexin V flow cytometry was performed using PI (ThermoFisher, Waltham, MA, USA, Catalog No. P1304MP) and Annexin V (ThermoFisher, Waltham, MA, USA, Catalog No. R37176). The flow cytometry data were collected using FACSDiva software (BD Biosciences, San Jose, CA, USA) and analyzed using FlowJo software (FlowJo LLC, Ashland, OR, USA).
Muralikrishnan V., Fang F., Given T.C., Podicheti R., Chtcherbinine M., Metcalfe T.X., Sriramkumar S., O’Hagan H.M., Hurley T.D, & Nephew K.P. (2022). A Novel ALDH1A1 Inhibitor Blocks Platinum-Induced Senescence and Stemness in Ovarian Cancer. Cancers, 14(14), 3437.
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4

CD133+ Cell Isolation from Tumor Tissue

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CD133 positive (CD133+) cells were isolated from freshly resected and physically/enzymatically dissociated tumor tissue samples using Magnetic-activated Cell Sorting (MACS) technique (Miltenyi Biotech, Bergisch Gladbach, Germany) and “EasySep Positive Selection Human PE Selection Kit (StemCell Technologies, (Vancouver, BC, Canada)” following the manufacturer’s protocol. Shortly, fresh tumor tissue samples were physically minced with a scalpel and exposed to enzymatic dissociation using 400 μg/ml Collagenase enzyme (GIBCO, New York, USA) at 37 °C for 3 h. Dissociated cells were filtered using a 70-μm cell strainer to get a single cell suspension. Cells were labeled with CD133/2-PE (Miltenyi Biotech clone AC133) antibody. After magnetic sorting, CD133 enriched (CD133+) and remaining (CD133−) cell populations from the same tissue samples were immediately washed and homogenized in “Lysis/Binding Buffer” of “mirVana miRNA Isolation Kit” (Ambion, Darmstadt, Germany) for further RNA isolation.
Karatas O.F., Suer I., Yuceturk B., Yilmaz M., Oz B., Guven G., Cansiz H., Creighton C.J., Ittmann M, & Ozen M. (2016). Identification of microRNA profile specific to cancer stem-like cells directly isolated from human larynx cancer specimens. BMC Cancer, 16, 853.
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5

Flow Cytometry Analysis of Cell Markers

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Flow cytometry analysis of surface markers was performed as previously described [43 (link)]. Intracellular staining was performed by using Fixation/Permeabilization Solution Kit (BD Pharmingen), where cells were incubated with primary antibody over night, followed by washing and then incubated with secondary antibody for 2 h. Cells stained with secondary antibodies alone were used as control for gating. The following antibodies were used: CD166-PE, CD9- FITC, CXCR4-PE, m/hCD44-APC (eBioscience, San Deigo, CA), CD133/2-PE and anti-m/hAPC-A2B5 (Miltenyi, Billerica, MA).
Joel M., Mughal A.A., Grieg Z., Murrell W., Palmero S., Mikkelsen B., Fjerdingstad H.B., Sandberg C.J., Behnan J., Glover J.C., Langmoen I.A, & Stangeland B. (2015). Targeting PBK/TOPK decreases growth and survival of glioma initiating cells in vitro and attenuates tumor growth in vivo. Molecular Cancer, 14, 121.
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6

Multiparameter Flow Cytometry Analysis

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We used the following antibodies: CD44-PE-Cy5 (1:100) (eBioscience), CD15-APC (1:5) (BD), CD133/2-PE (1:50) (Miltenyi Biotec). Annexin V-FITC Apoptosis Detection Kit was obtained from eBioscience. Staining was quantified by flow cytometry (Cytek DxP10).
Li Q., Lin H., Rauch J., Deleyrolle L.P., Reynolds B.A., Viljoen H.J., Zhang C., Zhang C., Gu L., Van Wyk E, & Lei Y. (2018). Scalable Culturing of Primary Human Glioblastoma Tumor-Initiating Cells with a Cell-Friendly Culture System. Scientific Reports, 8, 3531.
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7

Immunophenotyping of Stem Cells

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For flow cytometry, cells were stained with the following antibodies from Miltenyi Biotec: CD133/2-PE (clone: 293C3), CD34-APC (clone: AC136) and CD45-VioBlue (clone: 5B1) according to the manufacturer’s protocol (Miltenyi Biotec). To determine the cell viability, propidium iodide was added to the samples just before measurement. Flow cytometric analysis was performed using MACSQuant Analyzer 10 (Miltenyi Biotec).
Diener Y., Jurk M., Kandil B., Choi Y.H., Wild S., Bissels U, & Bosio A. (2015). RNA-based, transient modulation of gene expression in human haematopoietic stem and progenitor cells. Scientific Reports, 5, 17184.
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8

Comprehensive Immunophenotyping of Cell Populations

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Following fluorochrome-conjugated antibodies were used for the evaluation of surface markers: CD44-PE, CD24-PE, CD26-PE, CD271-PE, CD133/2-PE (Miltenyi Biotec, Germany); CD166-PE (ALCAM; Immunotech, France); CD274 (PD-L1; Sony Biotechnology, USA); CD44v6-PE (RD Systems, USA); CD184 (CXCR4-PE; eBioscience, USA). Dead cells were excluded from the analysis based on DAPI staining. Cells were analyzed using BD FACSCanto™ II flow cytometer (Beckton Dickinson, USA) equipped with FacsDiva program. FCS Express software was used for the evaluation.
Durinikova E., Kozovska Z., Poturnajova M., Plava J., Cierna Z., Babelova A., Bohovic R., Schmidtova S., Tomas M., Kucerova L, & Matuskova M. (2018). ALDH1A3 upregulation and spontaneous metastasis formation is associated with acquired chemoresistance in colorectal cancer cells. BMC Cancer, 18, 848.
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