The largest database of trusted experimental protocols

Fcs5 express software

Manufactured by De Novo Software
Sourced in United States

FCS5 express software is a data analysis tool for flow cytometry data. It provides users with capabilities to import, visualize, and analyze flow cytometry data files.

Automatically generated - may contain errors

4 protocols using fcs5 express software

1

Flow Cytometric Analysis of P-gp Expression

Check if the same lab product or an alternative is used in the 5 most similar protocols
Flow cytometric analysis was carried out on leukemic cells and on cell suspensions (106 cells/mL) of colon carcinoma cells obtained by incubating monolayer cell cultures with EDTA and trypsin. For the P-gp determination on the cell surface, the cells were incubated for 30 min at RT with MAb FITC conjugate CD243. After washing with ice-cold phosphate buffer saline (PBS, pH 7.2) with 1% bovine serum albumin (BSA, Sigma), cells were immediately analyzed. Dead cells were excluded from the analysis by gate on cells size before the acquisitions. Ten thousand events were acquired and fluorescence was analyzed with a FACScanto flow cytometer (Becton Dickinson) equipped with a 15 mW, 488 nm, air-cooled argon ion laser. The fluorescence emissions were collected through a 530 nm band-pass filter for fluorescein.
For quantitative evaluation of P-gp expression, the mean fluorescence channel (MFC) was calculated by FCS5 express software (De Novo software). P-gp content in arbitrary units was expressed as the ratio of the MFC of labeled resistant cells on the MFC of parental ones.
+ Open protocol
+ Expand
2

Evaluating Immune Phenotype and Docetaxel Response in Prostate Cancer

Check if the same lab product or an alternative is used in the 5 most similar protocols
To evaluate the immune phenotype, PC3 and PC3RR cells were detached with 0.25% trypsin EDTA (Sigma), washed with PBS and incubated with bv421-conjugated Integrin alpha-2 antibody (BD Bioscience, San Jose, CA, USA) in PBS/0.1% BSA (Sigma) for 30 min on ice prior to flow cytometric analysis. Propidium iodide (PI) solution (Sigma-Aldrich P4864) was added to exclude dead cells. After washing, cells were assayed using a CyAn ADP flow cytometer (Beckman Coulter, Brea, CA, USA) and data were analyzed using FCS5 Express Software (De Novo Software, Glendale, CA, USA). For cell cycle analysis, PCa and PCaRR cells were fixed with 70% ethanol, washed three times with PBS and stained for 3 h at room temperature with PI, then analyzed using flow cytometry.
To assess the effects of Docetaxel treatment, PC3, PC3RR, DU-145 and DU-145RR cells were seeded 9 × 104/well and 6 × 104/well, respectively, in 12-well plates, then treated with different concentrations (from 10 nM to 100 nM) of Docetaxel (Sigma-Aldrich) for 24 and 48 h. Then, apoptosis/necrosis were assayed by using PI/Annexin Pacific Blue staining (Thermo Fisher Scientific, Rockford, IL, USA) and evaluated using flow cytometry.
+ Open protocol
+ Expand
3

CD44v9 Expression and Cell Cycle Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cells were detached and incubated with APC-conjugated CD44 antibody (BD-PharMingen, San Diego, CA, USA 559942) or isotype control APC-conjugated IgG2B (BD-PharMingen) in PBS BSA 1% (Sigma) for 30 minutes on ice prior to flow cytometric analysis. Sytox Blue Stain (Life Technologies, Eugene, OR, USA S34857) was added to exclude dead cells. For CD44v8-10 staining 1×105 cells were incubated with 3 μg/ml anti-human CD44v9 primary antibody (clone: RV3) (Cosmo Bio Co. Ltd, Tokyo, Japan LKG-M001) in PBS 0.2 % BSA for 45 minutes at 4°C. Cells were then incubated with the secondary antibody APC-labeled Goat anti-Rat IgG (H+L) (Invitrogen, Carlsbad, CA, USA A10540) for 30 minutes at 4°C. After washing, cells were assayed using a CyAn ADP flow cytometer (Beckman Coulter, Brea, CA, USA) and data analyzed employing FCS5 express Software (De Novo Software). For cell cycle analysis, after labeling for CD44v9, the cells were fixed with 70% ethanol, a few drops at a time mixing the cells, and incubated on ice for 30 minutes. The cells were centrifuged at 500 x g for 10 minutes, washed once in PBS by centrifugation and resuspended in 1 ml PBS containing 5 μg/ml of propidium iodide. Samples were assayed by CyAn ADP flow cytometer and analyzed employing FCS5 express Software.
+ Open protocol
+ Expand
4

Measurement of Cellular Reactive Oxygen Species

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cells were detached with 0.25% trypsin/EDTA (Sigma), washed with PBS 0.1% BSA, and incubated with 200 μM H2O2 at 37 °C for 30 min. After 2 washes in PBS 0.1% BSA, the cells have been incubated with the cell permeant reagent 2′,7′-dichlorofluorescindiacetate (DCFDA) (Sigma), a fluorogenic dye that measures hydroxyl, peroxyl, and other ROS activity within the cell. After diffusion into the cell, DCFDA is deacetylated by cellular esterases to a non-fluorescent compound, which is later oxidized by ROS into 2′,7′-dichlorofluorescein (DCF). DCF is a highly fluorescent compound, and it has been detected by flow cytometry analysis using CyAn ADP flow cytometer (Beckman Coulter`), and data were analyzed with FCS5 Express software (De Novo Software).
+ Open protocol
+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required

Sign up now

Revolutionizing how scientists
search and build protocols!