Cellquest pro

Manufactured by BD

Sourced in United States, Germany, United Kingdom, France, Italy, Spain

CellQuest Pro is a software application for flow cytometry data acquisition and analysis. It provides a user-friendly interface for controlling flow cytometers and managing experimental data.

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Lab products found in correlation

Facscalibur, by BD (311 mentions) Facscalibur flow cytometer, by BD (109 mentions) Propidium iodide, by Merck Group (39 mentions) Facscalibur cytometer, by BD (37 mentions) Facscan, by BD (31 mentions) Facscan flow cytometer, by BD (17 mentions) Fitc annexin 5 apoptosis detection kit, by BD (13 mentions) Facscalibur system, by BD (13 mentions) Cba software, by BD (12 mentions) Facscanto 2, by BD (11 mentions) Facsaria 2, by BD (10 mentions) Facsdiva, by BD (10 mentions) Rnase a, by Merck Group (10 mentions) Facs caliber, by BD (10 mentions) Cellquest pro software, by BD (8 mentions) Propidium iodide, by BD (8 mentions) Lsrfortessa, by BD (7 mentions) Annexin 5 fitc, by BD (7 mentions) Facscanto, by BD (6 mentions) Aldefluor assay, by STEMCELL (6 mentions)

821 protocols using cellquest pro

Assessing Cell Viability and P2X7 Uptake

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Cell viability was assessed by annexin V/propidium iodide staining (eBioscience, San Diego, CA, USA). Briefly, cells were harvested with trypsin-EDTA, suspended in staining buffer, stained for 20 min with annexin V and propidium iodide, and analyzed using a BD FACSCalibur TM and BD CellQuest TM Pro (Becton, Dickinson Biosciences, San Jose, CA, USA).
P2X 7 channel uptake YO-PRO-1, a 629-Da fluorescent nucleic acid dye that is transported across the cell membrane by P2X 7 , was used to quantify the uptake of extracellular ATP, as previously described (Cankurtaran-Sayar et al. 2009) . In brief, cells were collected with trypsin-EDTA, centrifuged at 1,500 rpm for 5 min, washed once with cold phosphatebuffered saline, and stained with 2 μM YO-PRO-1 (Invitrogen, Carlsbad, CA, USA) for 30 min in the dark. The mean fluorescence intensity was then analyzed using a BD FACSCalibur TM and BD CellQuest TM Pro (Becton, Dickinson Biosciences, San Jose, CA, USA).

Ho V.M., Hirohashi N., Kong W.S., Yun G., Ota K., Itai J., Yamaga S., Suzuki K., Tanigawa K., Kanno M, & Shime N. (2018). Sera from Septic Patients Contain the Inhibiting Activity of the Extracellular ATP-Dependent Inflammasome Pathway. The Tohoku journal of experimental medicine, 245(3).

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Quantifying Apoptosis by Flow Cytometry

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Flow cytometry was performed using the BD FACSCalibur system (Fluorescence-activated cell sorter flow cytometer, BD, Heidelberg, Germany). Running samples with BD CellQuest Pro software (both from Becton, Dickinson and Company, New Jersey, USA), the number of events counted by the flow cytometer were at least 10000 per sample. All experiments were performed in triplicate. Co-staining for Annexin V and propidium iodide, respectively, was conducted using the Annexin-V-FLUOS Staining Kit (Roche Diagnostics, Basel, Switzerland), according to the manufacturer's instructions. Annexin V-positive and propidium iodide-negative cells were regarded as early apoptotic cells, while cells displaying Annexin V/propidium iodide co-staining represented the late apoptosis/necrotic cell fraction, respectively. Data analysis was done by performing quadrant statistics using BD CellQuest Pro software (BD, Heidelberg, Germany).

Eder S., Lamkowski A., Priller M., Port M, & Steinestel K. (2015). Radiosensitization and downregulation of heterogeneous nuclear ribonucleoprotein K (hnRNP K) upon inhibition of mitogen/extracellular signal-regulated kinase (MEK) in malignant melanoma cells. Oncotarget, 6(19), 17178-17191.

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Flow Cytometry Bacterial Cell Enumeration

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Flow cytometric measurements were carried out as described previously (Liu et al., 2017a (link)). One milliliter of water sample was stained with 10 μL/mL SYBR Green I (1:100 dilutions in dimethyl sulfoxide as the working solution; Invitrogen, United States), and incubated in the dark for 15 min at room temperature. FCM then was performed at a flow rate of 60 μL/min using a BD FACSCalibur system (BD Biosciences, San Jose, CA, United States) equipped with a 15-mW air-cooled argon laser, emitting at a fixed wavelength of 488 nm. All data were analyzed with the BD CellQuest Pro software (BD Biosciences). Green fluorescence was collected in the FL1 channel (520 ± 20 nm) and bacterial cell density was counted through the gating on a two-parameter dot-plot of green fluorescence against side scatter. Milli-Q cell-free water was used as the blank samples and sheath fluid as well in FCM. Positive signals of bacterial cell was separated from instrument noise or sample background by electronic gating with BD CellQuest Pro software (BD Biosciences). All samples were collected as logarithmic signals and were triggered on the green fluorescence channel. The coefficient of variation on FCM measurement for replicates should be below 2%, and all samples were measured in triplicate.

Liu J., Zhao R., Zhang J., Zhang G., Yu K., Li X, & Li B. (2018). Occurrence and Fate of Ultramicrobacteria in a Full-Scale Drinking Water Treatment Plant. Frontiers in Microbiology, 9, 2922.

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Intracellular ROS Production Quantification

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T24 cells were seeded into 6-well plates at a density of 5×10⁴ cells/well and cultured at 37°C with 5% CO₂ for 24 h. Cells were pretreated with N-acetyl-L-cysteine (NAC, Sigma-Aldrich; Merck KGaA) for 3 h, and treated with cisplatin (20 µM) and/or β-ELE (50 µg/ml) for another 12 h at 37°C, before being stained with DCFH-DA probe for 20 min at 37°C using the Reactive Oxygen Species Assay kit (Beyotime Institute of Biotechnology) according to the manufacturer's instructions. Intracellular production of ROS was evaluated by flow cytometry using CellQuest™ Pro software (version 5.1; BD CellQuest Pro; BD Biosciences,).

Gan D., He W., Yin H, & Gou X. (2019). β-elemene enhances cisplatin-induced apoptosis in bladder cancer cells through the ROS-AMPK signaling pathway. Oncology Letters, 19(1), 291-300.

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Flow Cytometric Analysis for Transfected Cell Selection

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For positive transfected cells selection, flow cytometric analysis was performed. Mouse anti-c-myc (9E10, in house) or mouse anti-NGFR (HB-8737, in house) antibodies were incubated with RBL-2H3 transfected cells or mock transfectants for 20 min at 4 °C. After washing with PBS, cells were stained with FITC-conjugated goat anti-mouse IgG (1:60, Sigma, Burlington, MA, USA) for 10 min at 4 °C and were analyzed by flow cytometry using a FACSCalibur (Becton Dickinson, Madrid, Spain) cytometer. A total of 10,000 events were acquired and data were analyzed with BD CellQuest Pro software (BD CellQuest Pro, BD Biosciences, Madrid, Spain).

Gómez-Melero S., García-Maceira F.I., García-Maceira T., Luna-Guerrero V., Montero-Peñalvo G., Caballero-Villarraso J., Túnez I, & Paz-Rojas E. (2022). Development of a High-Throughput Calcium Mobilization Assay for CCR6 Receptor Coupled to Hydrolase Activity Readout. Biomedicines, 10(2), 422.

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Apoptosis and CD69 Analysis of Th1/Th2 Cells

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To study the apoptosis of transfected Th1 and Th2 polarized cells, cells were transfected, rested for 20–24 h and cultured for 24 h or 48 h as described earlier in the Cell culture and transfections-section. 0.5×10⁶ cells per sample were then harvested, washed twice with PBS and once with 1xBinding Buffer ((5 mM HEPES, 70 mM NaCl, 2.5 mM CaCl₂, pH 7.4) in 2%FCS/PBS (w/v), 0.01% NaN₃). Cells were stained with Annexin V-FITC (BD Pharmingen, San Jose, CA) and incubated at RT for 20 min. Cells were then washed twice with Binding buffer. 20 s prior to analysis with FACSCalibur system, propidium iodide (PI; BD Pharmingen) was added to the sample. The data was analyzed with CellQuest Pro (BD Biosciences) or FlowJo (TreeStar Inc).
For CD69 analysis, transfected cells were cultured for 24 h in Th1 or Th2 polarizing conditions and 0.5×10⁶ cells per sample were harvested for staining. Cells were washed with 2% FCS/PBS, 0.01% NaN₃ and stained with CD69-FITC (BD Biosciences) or isotype control anti-mouse IgG1-FITC (MG101, Invitrogen). Cells were analyzed with the FACSCalibur system and analyzed with CellQuest Pro (both from BD Biosciences).

Kyläniemi M.K., Kaukonen R., Myllyviita J., Rasool O, & Lahesmaa R. (2014). The Regulation and Role of c-FLIP in Human Th Cell Differentiation. PLoS ONE, 9(7), e102022.

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Complement Binding Assay for Cells

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Cells used in FACS analysis were detached with 1 mM EDTA in PBS for 10 min and pre-incubated 10 min with 2.4G2 monoclonal antibody at 4 °C to block Fc receptors. Cells or S. pneumoniae were incubated with 3–5% mouse or human serum in 200 μL TC buffer (10 mM Tris-HCl, 140 mM NaCl, 2 mM CaCl₂, 2 mM MgCl₂, and 1% bovine serum albumin [BSA]), and complement binding to cells or organisms was detected by immunostaining with respective antibodies for 30 min at 4 °C. Cytometric analysis was performed using a FACScan (Becton Dickinson, San Jose, CA, USA) and CellQuestPro (BD Biosciences). Subsequent data analysis was performed with CellQuestPro (BD Biosciences).

Yang S.W., Park J.Y., Choi H., Yun T.J., Choi W.S., Kim M.K., Lee Y.K., Park M., Jin Y., Joo J.S., Choi I.S., Park S.H., Hwang H.S, & Kang Y.S. (2019). Dominant role of splenic marginal zone lipid rafts in the classical complement pathway against S. pneumoniae. Cell Death Discovery, 5, 133.

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Intracellular ROS Assessment in PBMC

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Intracellular ROS generation in PBMC cells (5 × 10⁵ pelleted cells) was measured by flow cytometric assay method [13 (link)] using a ROS-sensitive cell-permeable dye 2′ 7′ dihydrodichlorofluorescein diacetate (2′ 7′ H2DCFDA). In this method, 2′ 7′ H₂DCFDA oxidized to highly fluorescent 2′ 7′-dichlorofluorescein (2′ 7′ DCF) presence of ROS in the PBMC. The PBMC exhibited an increased fluorescence of oxidized DCF, as measured by a flow cytometer (FACSCalibur, Becton Dickinson, San Jose, CA) fitted with an argon-ion laser (15 mW) set to a wavelength of 488 nm. The fluorescence of DCF was collected in FL1 channel, equipped with a 530/30 nm band-pass filter. Fluorescence was measured in the long mode using “Cell Quest Pro” software (BD Bioscience, San Jose, CA) and expressed as geometrical mean fluorescence channel (GMFC). Cells were gated on the basis of their characteristic morphology, i.e., forward scatter and side scatter of monocytes and lymphocytes. Acquisitions were performed on 10000 gated events; while data analysis was carried out with “Cell Quest Pro” software (BD Bioscience).

Pramanik S., Banerjee K, & Mondal L.K. (2022). The Amelioration of Detrimental Biochemical Anomalies by Supplementing B, C, and E Vitamins in Subjects with Type 2 Diabetes Mellitus May Reduce the Rate of Development of Diabetic Retinopathy. Journal of Diabetes Research, 2022, 3886710.

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Cell Cycle Analysis by Flow Cytometry

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After the treatment of the cells with the indicated compound for 24 or 48 h, the cells were harvested by trypsinization, fixed with 70% (v/v) alcohol at −20 °C for 30 min and washed with PBS. The cells were centrifuged and re-suspended with 0.5 mL PI solution containing Triton X-100 (0.1% v/v), RNase (100 μg/mL), and PI (80 μg/mL). DNA content was detected using flow cytometric analysis (FACSan FL2 channel and analyzed with BD CellQuest^TM Pro software (Becton Dickinson

Chang Y.C., Tseng Y.L., Leu W.J., Du C.M., Jiang Y.H., Hsu L.C., Hsu J.L., Hou D.R, & Guh J.H. (2020). Discovery of Novel Agents on Spindle Assembly Checkpoint to Sensitize Vinorelbine-Induced Mitotic Cell Death against Human Non-Small Cell Lung Cancers. International Journal of Molecular Sciences, 21(16), 5608.

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EPC Surface Marker Expression Analysis

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Flavonoid treated EPCs were harvested and incubated with Fc Receptor (FcR) blocker at room temperature for 15 min. The EPCs then were incubated with each of the following fluorochrome conjugated antibodies: CD133 PE (Miltenyi Biotec, Germany), and VEGFR2/KDR/Flk-1 PE. Expression for each surface marker was analyzed using a FACS Calibur flowcytometry. Calculation of the percentage of markers was done based on percentage reduction of markers with the isotype using BD Cell QuestTM Pro software (Becton Dickinson, USA) [14, 15] .

Widowati W., Gunanegara R.F., Wargasetia T.L., Kusuma H.S.W., Arumwardana S., Wahyuni C.D., Girsang E., Lister I.N.E., Ginting C.N., Rizal R., Bachtiar I., Murti H., & Kim Y.H. (2021). EFFECT OF FLAVONOIDS ON OXIDATIVE STRESS, APOPTOSIS, AND CELL MARKERS OF PERIPHERAL BLOOD-DERIVED ENDOTHELIAL PROGENITOR CELLS: AN IN VITRO STUDY. International Journal of Applied Pharmaceutics.

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