PMDV-coated and uncoated Si particles were suspended at a concentration of 1 million per 100 μL blocking buffer PBS/1% BSA. APC-conjugated primary antibodies were added in the blocking buffer and incubated for 30 min at room temperature. Following three washes with 1 mL of PBS, fluorescence measurements were collected using a Guava flow cytometer (EMD, Billerica, MA, USA). Data were analyzed using the Flow Express software (De Novo Software, Los Angeles, CA, USA). For fluorescence microscopy detection, stained particles were first immobilized on poly-lysine coated glass slides. Images were acquired in an upright Olympus BX-50 microscope.
Flow express software
Manufactured by De Novo Software
Sourced in United States
Flow Express software is a data analysis tool designed for flow cytometry applications. It provides functionalities for data acquisition, visualization, and analysis of flow cytometry data.
Automatically generated - may contain errors
Lab products found in correlation
Gibco cell dissociation buffer, by Thermo Fisher Scientific (3 mentions)
Bx50 microscope, by Olympus (1 mentions)
Apoptosis detection kit, by Keygen Biotech (1 mentions)
Guava easycyte flow cytometry, by Merck Group (1 mentions)
Accuri c6, by BD (1 mentions)
Accuri c6 flow cytometer, by BD (1 mentions)
5 protocols using flow express software
1
Quantification of Surface Bound Antibodies
2
Apoptosis Detection in Cultured Cells
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The effect of apoptosis-induction was confirmed using the Apoptosis Detection Kit (Keygen, Nanjing, Jiangsu, China), according to the manufacturer’s protocol. Briefly, cells for transfection were detached with enzyme-free EDTA, washed twice with PBS and suspended at a concentration of 5×105 cells in 300 μL binding buffer. Annexin V-APC and 7-AAD were added to the cells and allowed to incubate for 15 min at room temperature in the dark. Apoptotic cells were identified using an Accuri C6 flow cytometer (BD Biosciences, San Jose, CA, United States) on the FL-4 and FL-3 channels. Data were analyzed using the Flow Express software (De Novo Software, Los Angeles, CA, United States).
3
Cell Surface Protein Analysis by Flow Cytometry
4
Cell Surface Marker Quantification
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Cells were detached with enzyme-free Gibco® Cell Dissociation Buffer (Invitrogen) and suspended at a concentration of 5 × 105 cells in 100 μL cold PBS/1% bovine serum albumin (BSA). Primary antibodies or corresponding isotype control antibodies were incubated with cells for 30 min on ice. Following two washes with 1 mL of PBS/1% BSA, fluorescence measurements were collected using an Accuri C6 flow cytometer (BD Biosciences, San Jose, CA, USA). Data were analyzed using the Flow Express software (De Novo Software, Los Angeles, CA, USA).
5
Quantifying Cell Adhesion Molecules
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E-selectin and ICAM-1 are molecules associated with cell-to-cell adhesion and are found on cell surfaces. The expression of these proteins was quantified using BD Accuri TM C6 Flow Cytometer (B.D. Biosciences). Cells were detached with enzyme-free Gibco cell dissociation buffer (Invitrogen) and suspended in cold PBS 200 ul of 1% bovine serum albumin (BSA) at a 1×106 cells concentration. The cells were treated with antibodies against each cell adhesion protein, P.E. Mouse Anti-Human CD62P targeting E-selectin and P.E. Mouse Anti-Human CD54 targeting ICAM-1, Anti-Human/Mouse beta-Catenin Alexa Fluor® 488, and Mouse Anti-Human CD144 were incubated with cells for 30 min on ice. Cells were then washed twice with 1 mL PBS/1% BSA, and fluorescence measurements were detected on an Accuri C6 flow cytometer. Analysis of the data was performed using the Flow Express software (De Novo Software, Los Angeles, CA, USA). The flow cytometry assay provides protein assay using live cells under different conditions.
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