Cells were dissociated with Accutase for 10 min at 37°C, triturated, and passed through a 40 μm cell strainer. Cells were then washed twice with flow cytometry buffer (BD Biosciences) and resuspended at a maximum concentration of 1 × 106 cells per 100 μL. Cells were stained with PE-Mouse anti-human CD44 or isotype control (PE-Mouse IgG1,κ, BD Biosciences, Cat# 555749) antibodies for 1 hr on ice. Cells were washed twice with flow cytometry buffer prior to analysis. Flow cytometry analysis was performed on an Accuri C6 flow cytometer (BD Biosciences). Flow cytometry files were analyzed using FlowJo (FlowJo LLC, Ashland, OR, USA).
Flow cytometry buffer
Manufactured by BD
Sourced in United States
Flow cytometry buffer is a specialized solution used in flow cytometry, a technique used to analyze and sort cells or other particles. The buffer is designed to maintain the physical and chemical properties of the samples being analyzed, ensuring accurate and reliable results. It helps to preserve the integrity of the cells or particles and facilitates their movement through the flow cytometer.
Automatically generated - may contain errors
Lab products found in correlation
Pe mouse igg1κ, by BD (2 mentions)
Accuri c6, by BD (2 mentions)
Accuri c6 flow cytometer, by BD (1 mentions)
Lsrii flow cytometer, by BD (1 mentions)
Mitosox red, by Thermo Fisher Scientific (1 mentions)
Antimycin a, by Merck Group (1 mentions)
Mitosox red, by Merck Group (1 mentions)
Attune nxt, by Thermo Fisher Scientific (1 mentions)
Rat isotype control igg, by BioLegend (1 mentions)
Facsaria 3, by BD (1 mentions)
Dmem 10 fbs, by Thermo Fisher Scientific (1 mentions)
Annexin 5 fitc and pi, by BD (1 mentions)
Custom matlab, by MathWorks (1 mentions)
Facsdiva software, by BD (1 mentions)
Facscalibur flow cytometer, by BD (1 mentions)
Cellquest software, by BD (1 mentions)
9 protocols using flow cytometry buffer
1
CD44 Surface Expression Analysis
2
Neonatal Hyperoxia Alters Mitochondrial ROS
3
Dissociation and Flow Cytometry Analysis
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Cells were dissociated with Accutase for 10 min at 37 °C, triturated, and passed through a 40-μm cell strainer. Cells were then washed twice with flow cytometry buffer (BD Biosciences) and resuspended at a maximum concentration of 5 × 106 cells per 100 μL. Flow cytometry analysis was performed on an Attune NxT (Thermo Fisher Scientific). Flow cytometry files were analyzed using with FlowJo (FlowJo LLC, Ashland, OR, USA).
4
CD44 Expression Analysis by Flow Cytometry
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Cells were dissociated with Accutase for 10 min at 37 °C, triturated, and passed through a 40 μm cell strainer. Cells were then washed twice with flow cytometry buffer (BD Biosciences) and resuspended at a maximum concentration of 1 × 106 cells per 100 μL. Cells were stained with PE-Mouse anti-human CD44 or isotype control (PE-Mouse IgG1,κ, BD Biosciences, Cat# 555749) antibodies for 1 h on ice. Cells were washed twice with flow cytometry buffer prior to analysis. Flow cytometry analysis was performed on an Accuri C6 flow cytometer (BD Biosciences). Flow cytometry files were analyzed using FlowJo (BD).
5
Single-cell Genetic Modification in HEK293
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HEK293 cells were transfected in 12-well tissue culture plates at 40% confluence with the following reagents per well: 600 ng pCMV-BE4-Gam, 200 ng pEF1α-BFP, 200 ng pMT-sgRNA, 1.5 μl Lipofectamine 3000 Transfection Reagent and 2 μl P3000 reagent. After 48 h, cells were dissociated with Accutase for 5 min at 37°C, triturated and passed through a 40 μm cell strainer. Cells were then washed twice with flow cytometry buffer (BD Biosciences) and resuspended at a maximum concentration of 5 × 106 cells per 100 μl. Single GFP+ cells were sorted into a single well of a 96-well plate and expanded to a 24-well plate prior to analysis.
6
CX3CR1 Expression on Prostate Cancer Cells
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PC3-ML human prostate carcinoma cells were grown in DMEM (+10% FBS; Gibco) to 80% confluency and were then trypsinized, washed, and resuspended at a concentration of 106 cells/ml MACS flow cytometry buffer (Miltenyi Biotec). MMDCs were resuspended at a concentration of 106 cells/ml MACS flow cytometry buffer. 105 cells were stained with allophycocyanin (APC)-conjugated rat anti–human CX3CR1 IgG (BioLegend) or rat isotype control IgG (BioLegend) for 60 min at 4°C. Cells were then washed with and resuspended in MACS flow cytometry buffer and subjected to flow cytometry on a FACS ARIA III (BD). The data were analyzed with the FlowJo software.
7
Macrophage Polarization and Cell Apoptosis
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After treatment or transfection, macrophages were collected and resuspended in a flow cytometry buffer (BD Biosciences, San Diego, CA, USA). Then, macrophages were stained with anti-CD11b (ab24874) plus anti-iNOS (M1 marker, ab283655) or anti-CD206 (M2 marker, ab270682). The rates of M1 cells (CD11b+ and iNOS+) and M2 cells (CD11b+ and CD206+) were analyzed by flow cytometer and FlowJo software.
For detecting apoptosis rate, co-cultured AC16 cells were collected and suspended with binding buffer, followed by stained with Annexin V-FITC and PI (BD Biosciences). Cell apoptosis rate was assessed under a flow cytometer.
For detecting apoptosis rate, co-cultured AC16 cells were collected and suspended with binding buffer, followed by stained with Annexin V-FITC and PI (BD Biosciences). Cell apoptosis rate was assessed under a flow cytometer.
8
Cell Dissociation and Flow Cytometry
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Cells were dissociated with Accutase for 10 min at 37°C, and passed through a 40 μm cell strainer. Cells were then washed twice with flow cytometry buffer (BD Biosciences) and resuspended at a maximum concentration of 5 × 106 cells per 100 μl. Flow cytometry analysis was performed on an ACCURI C6 (BD Biosciences). Flow cytometry sorting was performed on a FACSAria IIu. Flow cytometry files were analyzed using with FACSDiva software (BD Biosciences), FlowJo (FlowJo LLC, Ashland, OR, USA), and custom Matlab (MathWorks, Natick, MA, USA) script.
9
Phenotypic Characterization of Rat BMSCs
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An analysis of cell surface molecules was performed on passage 3 cultures of rat BMSCs by flow cytometry with the following procedure: the medium was removed from the flasks and the cell layers were then washed twice with PBS and detached with 0.05% trypsin-EDTA (HyClone, Logan, UT, USA) for 2 min at room temperature. The BMSCs were collected by centrifugation (179 x g, at room temperature) and washed in flow cytometry buffer (BD Biosciences, Franklin Lakes, NJ, USA) consisting of 2% bovine serum albumin and 0.1% sodium azide in PBS. Subsequently, the cells were incubated with fluorescein-5-isothiocyanate (FITC)-conjugated monoclonal antibodies against CD44 (553133), CD45 (561867) and CD90 (561969), and phycoerythrin (PE)-conjugated antibody against CD34 (550619) (all from BD Biosciences). Following incubation with the antibodies for 30 min at 4˚C, the cells were washed with flow cytometry buffer. The washed cells were pelleted and resuspended in flow cytometry buffer containing 1% paraformaldehyde for 20 min. Non-specific fluorescence was determined using equal aliquots of the cell preparation that were incubated with anti-mouse monoclonal antibodies. Data were acquired and analyzed on a FACSCalibur flow cytometer with CellQuest software (BD Biosciences).
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