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Cellquest pro 6

Manufactured by BD
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CellQuest Pro 6.0 software is a data acquisition and analysis software designed for flow cytometry applications. The software enables users to collect, analyze, and manage flow cytometry data.

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Lab products found in correlation

Facscalibur flow cytometer, by BD (12 mentions) Facscalibur, by BD (8 mentions) Facscalibur cytometer, by BD (2 mentions) 7 aminoactinomycin d, by BD (2 mentions) Pe conjugated anti cd133, by BD (1 mentions) Propidium iodide, by Merck Group (1 mentions) Brdu elisa kit, by Roche (1 mentions) Pharmingen pi rnase staining buffer, by BD (1 mentions) Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions) Pi rnase staining buffer, by BD (1 mentions) Fcs express v3, by De Novo Software (1 mentions) Modfit lt software, by BD (1 mentions) Monensin, by Merck Group (1 mentions) Ionomycin, by Merck Group (1 mentions) Propidium iodide solution, by Thermo Fisher Scientific (1 mentions) Dispase, by Thermo Fisher Scientific (1 mentions) Collagenase type 4, by Thermo Fisher Scientific (1 mentions) Cacl2, by Merck Group (1 mentions) Cytofix cytoperm solution, by BD (1 mentions) Lsrfortessa flow cytometer, by BD (1 mentions)

Close spelling variants of this product

CellQuest Pro Software 6.0.4 BD Cellquest Pro software version 6.0 BD CellQuest Pro (version 6.0) CellQuest Pro CellQuest 6.0 CellQuest

28 protocols using cellquest pro 6

1

Evaluating CD133, CD44, and CD24 Expression

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CD133 surface expression was evaluated in both MCF10A and MCF10DCIS cells by means of flow cytometry after direct staining with phycoerythrin (PE)‐conjugated anti‐CD133/2 monoclonal antibody (293C3, Miltenyi Biotec, Bologna, I), as previously reported.8The expression of CD44 and CD24 surface antigens were evaluated following a previously described procedure.14 Briefly, in a one‐tube assay, cells were stained with allophycocyanin (APC)‐conjugated anti‐CD44 and fluorescein isothiocyanate (FITC)‐conjugated anti‐CD24 monoclonal antibodies (Becton Dickinson, San José, CA).
All samples were analyzed by a FACSCalibur flow cytometer (Becton‐Dickinson) with CellQuest Pro 6.0 software (Becton‐Dickinson). Data collected from 10 000 cells are shown as percentage of positive cells or as mean fluorescence intensity (MFI) values.
Bertagnolo V., Grassilli S., Volinia S., Al‐Qassab Y., Brugnoli F., Vezzali F., Lambertini E., Palomba M., Piubello Q., Orvieto E., Natali C., Piva M.R., Croce C.M, & Capitani S. (2019). Ectopic expression of PLC‐β2 in non‐invasive breast tumor cells plays a protective role against malignant progression and is correlated with the deregulation of miR‐146a. Molecular Carcinogenesis, 58(5), 708-721.
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2

Flow Cytometry for Stem Cell Markers

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The expression of CD133, EpCAM and CD44 surface antigens were evaluated by flow cytometry following a previously described procedure [21 (link)]. In a one-tube assay, cells were stained with phycoerythrin (PE)-conjugated anti-CD133/2 (293C3) and fluorescein isothiocyanate (FITC)-conjugated anti-CD326 (EpCAM) (Miltenyi Biotec, Bologna, I) or with PE-conjugated anti-CD133, FITC-conjugated anti-EpCAM and allophycocyanin (APC)-conjugated anti-CD44 (Becton Dickinson, San José, CA) monoclonal antibodies. All samples were analyzed by a FACSCalibur flow cytometer (Becton Dickinson) with the CellQuest Pro 6.0 software (Becton Dickinson). 20,000 non-debris events in the morphological gate were recorded for each sample. All antibodies were titrated under assay conditions and optimal photomultiplier (PMT) gains were established for each channel, as previously reported [22 (link)].
Data were analysed using FlowJo™ software (TreeStar, Ashland, OR) and reported as percentage of positive cells or as mean fluorescence intensity (MFI) values.
Brugnoli F., Grassilli S., Lanuti P., Marchisio M., Al-Qassab Y., Vezzali F., Capitani S, & Bertagnolo V. (2017). Up-modulation of PLC-β2 reduces the number and malignancy of triple-negative breast tumor cells with a CD133+/EpCAM+ phenotype: a promising target for preventing progression of TNBC. BMC Cancer, 17, 617.
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3

Characterization of Periosteal Cell Cultures

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Single cell suspensions of clonal periosteal cell cultures with different differentiation potential were washed in PBS/0.5%BSA [11 (link)]. Cells were incubated with titrated primary staining reagents for 15 min on ice (2.5 x 105 cells/0.1ml in PBS/0.5%BSA). Fluorescein isothiocyanate (FITC) labelled mouse anti-human CD105 (endoglin; SH-2) was purchased by Acris Antibodies (Acris Antibodies, Hiddenhausen, Germany). FITC labeled mouse anti-human CD44, CD45, and CD90 (Thy-1), and R-Phycoerythrin (PE) labeled mouse anti-human CD14, CD34, CD73 (SH-3), and CD166 (ALCAM) were purchased from Pharmingen (Heidelberg, Germany). Prior to the analysis in a FACS-Calibur cytometer (Becton Dickinson, Heidelberg, Germany), dead cells and debris were stained with propidium iodide PI (Sigma) and excluded. Data were evaluated using CellQuest Pro 6.0 software (Becton Dickinson).
Stich S., Loch A., Park S.J., Häupl T., Ringe J, & Sittinger M. (2017). Characterization of single cell derived cultures of periosteal progenitor cells to ensure the cell quality for clinical application. PLoS ONE, 12(5), e0178560.
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4

EpCAM Expression Quantification in hBTSCs

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The expression of EpCAM in hBTSC was evaluated by flow cytometry by direct staining with a fluorescein isothiocyanate (FITC)-conjugated anti-CD326 (EpCAM) monoclonal antibody (Miltenyi Biotec, Bologna, I), as suggested by manufacturer’s instructions. To assess the role of Vav1 on EpCAM levels, this surface antigen was evaluated in hBTSCs subjected to silencing of Vav1 with specific siRNAs during the first or the last 7 days of pancreatic differentiation. Non-specific fluorescence was assessed by using an isotype-matched control, Mouse IgG1-PE (Immunotech, Coulter Company, Marseille, F) and 7-amino-actinomycin D (Becton Dickinson, San Josè, CA) was added to the samples to exclude dead cells. All the samples were then analyzed by a FACSCalibur flow cytometer (Becton Dickinson) with the CellQuest Pro 6.0 software (Becton Dickinson). Data collected from 10,000 cells are shown as a percentage of positive cells.
Brugnoli F., Grassilli S., Cardinale V., Carpino G., Gaudio E., Alvaro D., Capitani S, & Bertagnolo V. (2020). Vav1 Sustains the In Vitro Differentiation of Normal and Tumor Precursors to Insulin Producing Cells Induced by all-Trans Retinoic Acid (ATRA). Stem Cell Reviews and Reports, 17(2), 673-684.
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5

Cell Viability, Proliferation, and Cycle Analysis

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Cell viability was determined using MTT metabolism test, as described [20 (link),33 (link)]. Cell proliferation was assessed using ELISA BrdU kit (Roche Diagnostics GmbH, Mannheim, Germany), according to the manufacturer’s protocol. For cell cycle analysis, cells were collected by trypsinization, fixed in 70% ethanol and stained with propidium iodide solution (3.8 mM sodium citrate, 50 mg/ml RNAse A, 500 mg/mL PI, in PBS). DNA content analyses were performed using FACS Calibur flow cytometer (BD Biosciences, Waltham, MA, USA) and the BD CellQuest Pro 6.0 software (BD Biosciences, San Jose, CA, USA). At least 10,000 events were analysed for each sample.
Król S.K., Kaczmarczyk A., Wojnicki K., Wojtas B., Gielniewski B., Grajkowska W., Kotulska K., Szczylik C., Czepko R., Banach M., Kaspera W., Szopa W., Marchel A., Czernicki T, & Kaminska B. (2020). Aberrantly Expressed RECQL4 Helicase Supports Proliferation and Drug Resistance of Human Glioma Cells and Glioma Stem Cells. Cancers, 12(10), 2919.
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6

Cell Cycle and Granularity Analysis by Flow Cytometry

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For cell cycle and cellular granularity analysis, cells were seeded at a density of 2 × 105 cells/well. Cell cycle analysis was performed by flow cytometry using BD Pharmingen PI/RNase Staining Buffer (BD Biosciences, USA). Briefly, cells were collected by trypsinization, fixed in 70% ethanol and stained in PI buffer (500 μL/1 × 106 cells). DNA content analyses were performed using FACScalibur flow cytometer (BD Biosciences, USA) and the BD CellQuest Pro 6.0 software (BD Biosciences, USA). At least 10,000 events were analysed for each sample.
Wojnicki K., Kaczmarczyk A., Wojtas B, & Kaminska B. (2023). BLM helicase overexpressed in human gliomas contributes to diverse responses of human glioma cells to chemotherapy. Cell Death Discovery, 9, 157.
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7

Flow Cytometry Analysis of CSC Markers

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HCT116 cells were stained with monoclonal antibodies (BD Biosciences, San Jose, CA, USA) characteristic for some CSC-specific antigens, namely, anti-CD29-APC (clone MAR4, IgG1κ) and anti-CD44-FITC (clone C26, IgG2bκ). In addition, we used an anti-CD133/2-PE (clone 293C3, IgG2bκ) monoclonal antibody from Miltenyi Biotec (Bergisch Gladbach, Germany). Cells were incubated for 30 min in the dark, washed, and resuspended with PBS containing 1 mM EDTA for a FACS analyses, which were performed using FACS Calibur flow cytometer (BD Biosciences, San Jose, CA, USA) and BD CellQuest Pro 6.0 software. The frequency of positive cells was compared to untreated control cells. We used unstained cells to set a threshold for the positive signal.
Szaryńska M., Olejniczak-Kęder A., Podpłońska K., Prahl A, & Iłowska E. (2023). Bradykinin and Neurotensin Analogues as Potential Compounds in Colon Cancer Therapy. International Journal of Molecular Sciences, 24(11), 9644.
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8

Analyzing p53 Transcriptional Activity

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LN18 cells (2.5 × 105 cells/well) were co-transfected with the plasmid carrying the gene encoding a wild-type p53 under the CMV promoter (pC53-SN3, 0.1 µg) [45 (link)] or empty vector (pcDNA3.1, 0.3 µg) and the plasmid encoding EGFP protein (pEGFP-N1, 0.3 µg) using Lipofectamine 2000 (Invitrogen, USA). Transfection efficiency and the p53 transcriptional activity were assessed with the p53-responsive luciferase reporter assay, based on two constructs: pPG13-Luc and pMG13-Luc, as described [24 ]. At 24 h after transfection cells were trypsinised, fixed and stained in 15 µM DRAQ5 in 4% PFA solution and a cell cycle was analysed using FACScalibur flow cytometer (BD Biosciences, USA) and the BD CellQuest Pro 6.0 software (BD Biosciences, USA). At least 10,000 events of GFP+ cells were analysed for each sample.
Wojnicki K., Kaczmarczyk A., Wojtas B, & Kaminska B. (2023). BLM helicase overexpressed in human gliomas contributes to diverse responses of human glioma cells to chemotherapy. Cell Death Discovery, 9, 157.
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9

Cell Cycle Analysis via Flow Cytometry

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DNA content distribution was quantified using the PI/RNase Staining Buffer (BD, Franklin Lakes, NJ, USA). Each cell was fixed with 70% ethanol, stained with PI, and then observed, with apoptotic cells being represented by the sub-G1 population. The fluorescence intensity for each cell was detected via flow cytometry using a FACSCalibur instrument (BD, Franklin Lakes, NJ, USA), and data were analyzed using BD CellQuest Pro 6.0 software (BD, Franklin Lakes, NJ, USA).
Choi H.S., Kim Y.K, & Yun P.Y. (2021). Cisplatin Plus Cetuximab Inhibits Cisplatin-Resistant Human Oral Squamous Cell Carcinoma Cell Migration and Proliferation but Does Not Enhance Apoptosis. International Journal of Molecular Sciences, 22(15), 8167.
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10

Murine Dendritic Cell HO-1 Expression

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Murine DCs were fixed in acetone, permeabilized, and stained with rabbit anti-HO-1 primary polyclonal antibody, followed by a FITC-conjugated goat anti-rabbit secondary mAb. Cells were analyzed using a FACSCalibur with Cellquest Pro 6.0 software (BD Biosciences, USA).
Zhao Y., Jia Y., Wang L., Chen S., Huang X., Xu B., Zhao G., Xiang Y., Yang J, & Chen G. (2018). Upregulation of Heme Oxygenase-1 Endues Immature Dendritic Cells With More Potent and Durable Immunoregulatory Properties and Promotes Engraftment in a Stringent Mouse Cardiac Allotransplant Model. Frontiers in Immunology, 9, 1515.
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