The largest database of trusted experimental protocols

Facs canto 2

Manufactured by Beckman Coulter
Sourced in United States

The FACS Canto II is a flow cytometry instrument designed for advanced multiparameter analysis. It incorporates a three-laser configuration and can detect up to 10 fluorescent parameters simultaneously. The system is capable of high-speed data acquisition and analysis for a wide range of applications in research and clinical laboratories.

Automatically generated - may contain errors

6 protocols using facs canto 2

1

Evaluating M1 and M2 Macrophage Markers

Check if the same lab product or an alternative is used in the 5 most similar protocols
The mean fluorescence intensity of M1 or M2 phenotype cells was determined by evaluating the level of their corresponding markers: CD86 and CD206, respectively, via flow cytometry assay. After treatment in the designed study groups, the BV-2 cells were collected and suspended in phosphate-buffered saline (PBS). Subsequently, the BV-2 cells were stained with the fluorescence-labeled CD86 (R-PE, BD Pharmingen, Franklin Lakes, NJ, USA, 09275B, 1:100) and CD206 (Alexa Fluor-647, BD Pharmingen, Franklin Lakes, NJ, USA, 565250, 1:200) antibodies for 30 min in the dark at 4 °C. FACS Canto II (Beckman Coulter, Indianapolis, IN, USA) was applied for analyzing the expression of CD86 and CD206.
+ Open protocol
+ Expand
2

Cytokine Profiling of Ag85A and ESAT-6 Responses

Check if the same lab product or an alternative is used in the 5 most similar protocols
Expression of IL-2, IFNγ and IL-17 by mononuclear cells in response to restimulation by Ag85A and ESAT-6 peptides was assayed by ICS, and data were analyzed using a hierarchical gating strategy, as outlined elsewhere [17 (link), 19 (link)]. Fluorochrome antibodies used for staining included CD3-APC-eFluor780 (Invitrogen), CD4-APC, CD8-PE-Cy7, IFNγ-PerCP Cy5.5, IL-17A-PE/Dazzle 594 (all from Biolegend) and IL-2-PE (BD Pharmingen). For tetramer staining, single cell suspensions of 1–2 × 106 cells per well were stained with the fluorescent antibodies CD3e-Pacific Blue and CD8-FITC (BD Pharmingen) and the PE-labeled MVPGGQSSF/H-2d tetramer (MHC Tetramer Production Facility, Baylor College of Medicine, Houston, Texas) for 45 minutes at room temperature. Cells were then washed twice with PBS, fixed, and 200,000 events were acquired on the FACS Canto II (Beckman Coulter).
+ Open protocol
+ Expand
3

Quantifying PD-L1 Secretion in CHO Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
CHO cells expressing either human PD-L1 or human CD80 fused to miRFP670 (table S3) were plated at 100,000 cells per well into a U-bottom 96-well plate and centrifuged at 300g for 5 min, and the supernatant was discarded. Two hundred–microliter supernatant containing secreted PD-L1 nanobody was added to CHO cell lines either neat or diluted in 1:10 and 1:100 in FACS buffer. Cells were incubated at 4°C for 30 min before being washed in FACS buffer for analysis on a Beckman Coulter FACSCanto II (Fig. 6F).
+ Open protocol
+ Expand
4

Macrophage Surface Marker Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols
Macrophages were incubated with TruStain FcX antibody (BioLegend, San Diego, CA, USA) for 10 min on ice to prevent unspecific binding of antibodies to Fc receptors. Then, cells were stained with rat anti-mouse CD86-APC (clone GL1, BioLegend, San Diego, CA, USA) or rat anti-mouse CD206-Brilliant Violet 421 (clone V068C2, BioLegend, San Diego, CA, USA) for 20 min at 4 °C in the dark and further washed with PBS supplemented with 2% FCS and 1 mM EDTA. Unstained cells were used as negative control for fluorescence. All data were collected by flow cytometry using FACSCanto II and DIVA software (Beckman Coulter, Brea, CA, USA).
+ Open protocol
+ Expand
5

Cytokine and Immunoglobulin Profiling in Chemotherapy

Check if the same lab product or an alternative is used in the 5 most similar protocols
Prior to treatment, and 2 weeks after the initial induction of chemotherapy, the serum levels of the cytokines IL-8, IL-6, IL-10, IL-1β, and sIL-2R were measured using the IMMULITE-1000 Immunoassay System (Siemens Healthineers, Erlangen, Germany) while the immunoglobulins IgA, IgG, IgM and IgE were detected by the BN II system (Siemens Healthineers). Lymphocyte subsets in the peripheral blood, including T, B, natural killer, cytokine-induced killer, T helper (Th), suppressor T (Ts), Th/Ts, and regulatory T cells (Treg) were tested using a FACSCanto™ II flow cytometer (Beckman Coulter, Inc., Brea, CA, USA), using a BD Multitest™ 6‐color TBNK kit and DIVA software.
+ Open protocol
+ Expand
6

Quantifying Cellular ROS Levels

Check if the same lab product or an alternative is used in the 5 most similar protocols
Alterations in the ROS levels were determined by dihydroethidium (DHE) staining (Molecular Probes, Eugene, OR, USA), which exhibits blue-fluorescence in the cytosol and can be oxidized by ROS in cells to form ethidium oxide, which can be incorporated into chromosomal DNA to produce red-fluorescent. Briefly, the cells were incubated with 4 μM DHE for 15 min at 37 °C. After washing twice with PBS, the population of ethidium bromide-stained cells was measured using FACS Canto II (EPICS XL, Beckman-Coulter, Miami, FL, USA and FACS Canto II, BD Biosciences, Mississauga, ON, USA) as the fraction of cells producing intracellular ROS, according to the manufacturer's protocol.
+ 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!