Facslyric system

Manufactured by BD

Sourced in United States

The BD FACSLyric system is a flow cytometry instrument designed for cell analysis and sorting. It features high-speed data acquisition and multi-parameter detection capabilities.

Automatically generated - may contain errors

Lab products found in correlation

Anti cd45ra pe cy7, by BD (2 mentions) Anti cd4 v500, by BD (2 mentions) Anti cxcr5 apc, by BioLegend (2 mentions) Anti foxp3 alexa fluor 488, by BD (2 mentions) Perm wash buffer, by BD (2 mentions) Transcription factor buffer, by BD (2 mentions) Anti cd3 v450, by BD (2 mentions) Anti pd 1 pe cy7, by BD (2 mentions) Alexa fluor 647 conjugated f ab 2 goat anti mouse igg h l secondary antibody, by Thermo Fisher Scientific (1 mentions) Fitc conjugated goat anti mouse igg, by Thermo Fisher Scientific (1 mentions) Vimentin, by Santa Cruz Biotechnology (1 mentions) Hoechst stain, by Thermo Fisher Scientific (1 mentions) Airyscan, by Zeiss (1 mentions) Click it edu cell proliferation kit, by Thermo Fisher Scientific (1 mentions) Panobinostat, by Selleck Chemicals (1 mentions) Xtt kit, by Roche (1 mentions) Adriamycin, by Selleck Chemicals (1 mentions) Rpmi 1640, by Thermo Fisher Scientific (1 mentions) Polybrene, by Merck Group (1 mentions) Phosphate buffered saline (pbs), by Lonza (1 mentions)

Close spelling variants of this product

BD FACSLyric (245 citations) BD FACSLyric Flow Cytometry System (6 citations) BD FACSLyric™ Research System (3 citations)

21 protocols using facslyric system

Feline PD-1 and PD-L1/PD-L2 Binding Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols

To examine whether feline PD-1 binds to PD-L1/PD-L2, 2 × 10⁵ fePD-1–EGFP- and fePD-L1–EGFP-expressing cells were incubated for 30 min with 10 μg/mL of either fePD-1–Ig, fePD-L1–Ig, or fePD-L2–Ig at room temperature (RT), followed by another 30 min incubation with Alexa Fluor 647-conjugated F(ab′)₂-goat anti-rabbit IgG (H+L) secondary antibody (Thermo Fisher Scientific). Rabbit IgG (Southern Biotech) was used as a control protein. Cell fluorescence was analyzed using BD FACSLyric system (BD Biosciences, Franklin Lakes, NJ, USA).
To examine PD-L1 expression on cell lines, 2 × 10⁵ cells were incubated for 30 min with 10 μg/mL CL1Mab-7 or mouse IgG₁κ isotype-matched control antibody (15H6; Southern Biotech) at RT, followed by another 30 min incubation with Alexa Fluor 647-conjugated F(ab′)₂-goat anti-mouse IgG (H+L) secondary antibody (Thermo Fisher Scientific). Cell fluorescence was analyzed using BD FACSLyric system (BD Biosciences). For fePD-1–EGFP- and fePD-L1–EGFP-expressing cells, only EGFP-positive cells were gated and subjected to further analysis.

Maekawa N., Konnai S., Asano Y., Otsuka T., Aoki E., Takeuchi H., Kato Y., Kaneko M.K., Yamada S., Kagawa Y., Nishimura M., Takagi S., Deguchi T., Ohta H., Nakagawa T., Suzuki Y., Okagawa T., Murata S, & Ohashi K. (2023). Molecular characterization of feline immune checkpoint molecules and establishment of PD-L1 immunohistochemistry for feline tumors. PLOS ONE, 18(1), e0281143.

+ Open protocol

+ Expand

Quantifying Apoptosis and Vimentin Expression

Check if the same lab product or an alternative is used in the 5 most similar protocols

SH-SY5Y or WJMSCs were washed twice with PBS then incubated with 5 μL FITC Annexin V and propidium iodide reagent for 15 min at RT in the dark according to the instruction. The apoptotic cells were analyzed using flow cytometry where 10,000 events were acquired in BD FACSLyric^™ system. For marker expression, WJMSCs with or without GA treatment were collected and fixed with 4% paraformaldehyde solution then washed twice with PBS. WJMSCs were then stained with Vimentin (Santa Cruz; sc-32322, 1:100) for 30 min and incubated with secondary FITC-conjugated goat anti-mouse IgG (Invitrogen; A11001, 1:100) in the dark for 30 min. The cells were acquired in BD FACSLyric^™ system.

Chen W.S., Lin T.Y., Kuo C.H., Hsieh D.J., Kuo W.W., Liao S.C., Kao H.C., Ju D.T., Lin Y.J, & Huang C.Y. (2023). Ginkgolide A improves the pleiotropic function and reinforces the neuroprotective effects by mesenchymal stem cell-derived exosomes in 6-OHDA-induced cell model of Parkinson’s disease. Aging (Albany NY), 15(5), 1358-1370.

+ Open protocol

+ Expand

Cellular Imaging and Flow Cytometry

Check if the same lab product or an alternative is used in the 5 most similar protocols

A total of 1 × 10⁶ cells were washed and fixed in 100% methanol for 10 min. Slides were prepared and stained with 5mg/mL Hoechst stain (Thermo Scientific, Rockford, IL, USA, Cat. No 62249) as per the manufacturer’s protocol. Slides were imaged at 40× in a standard DAPI filter set using a Carl Zeiss LSM880 confocal microscope with Airyscan (Peabody, MA, USA).
For Syto^TM 16 staining, 5 × 10⁵ cells were washed with 1X PBS containing 1% BSA and were stained with SYTO^TM 16 Green Fluorescent Nucleic Acid Stain (Life Technologies, Oregon, OR, USA, Cat no. S7578) as per manufacturer’s instructions. Stained cells were analyzed on BD FACSLyric^TM system (San Jose, CA, USA).

Katiyar S., Shah A., Rahman K., Tripathy N.K., Kashyap R., Nityanand S, & Chaturvedi C.P. (2023). Analysis of Immunophenotypic Changes during Ex Vivo Human Erythropoiesis and Its Application in the Study of Normal and Defective Erythropoiesis. Cells, 12(9), 1303.

+ Open protocol

+ Expand

Multicolor Flow Cytometry Immunophenotyping

Check if the same lab product or an alternative is used in the 5 most similar protocols

A total of 2 × 10⁵ cells were harvested on an alternate day and washed with a FACS buffer containing 1X PBS, 5 mM MgCl₂, and 1% BSA. Cells were stained with a cocktail of antibodies for each panel as mentioned in Table 1, Table 2 and Table 3 as per the manufacturer’s recommendation in a staining buffer containing 1X PBS, 0.09% BSA, 5 mM MgCl₂, 50 ug/mL DNAse I, and 5 μL of FCR blocking reagent. They were incubated for 30 min at room temperature. The cells were washed and resuspended in FACS buffer and were analyzed on a BD FACSLyric^TM system (San Jose, CA, USA). BD FACSuite^TM software (version 1.5) was used for gating and analysis.

+ Open protocol

+ Expand

Incorporating Cy5-eGFP mRNA into Extracellular Vesicles

Check if the same lab product or an alternative is used in the 5 most similar protocols

To incorporate Cy5‐eGFP mRNA in EVs, the HTB‐177 cells (1 × 10⁶ cells/T75 flask) were cultured in 15 mL RMPI‐1640 media. The conditioned media was replaced with fresh media after 24 h, and cells were incubated for a period of 24 h. Then next day, 100 µL of LNPs containing 100 µg Cy5 eGFP mRNA were added to HTB cells and incubated at 37 °C for 24 h. EVs were isolated/pre‐enriched by ultracentrifuge method as described above. CD63⁺ EVs were captured by magnetic dynabeads conjugated anti‐CD63 antibody (Immunoaffinity assay: Exosome‐Human CD63 isolation/detection reagent for cell culture medium, Thermo Fisher Scientific, cat.#: 10606D). In the binding reaction, 30 µL of CD63‐antibody conjugated beads were incubated with 60 µg of EVs. CD63⁺ EVs captured/immobilized by CD63‐antibody were acquired on a BD FACSLyric system (BD Biosciences) to detect Cy5‐eGFP mRNA. The data were analyzed using FlowJo software (TreeStar Inc.). The EVs from untreated cells were used as controls. For negative controls, 30 µL of CD63‐antibody conjugated beads alone (without EVs or LNPs) were incubated with an equivalent volume of PBS. Additionally, LNPs were also used as control to examine the contamination factor of LNPs.

Nawaz M., Heydarkhan‐Hagvall S., Tangruksa B., González‐King Garibotti H., Jing Y., Maugeri M., Kohl F., Hultin L., Reyahi A., Camponeschi A., Kull B., Christoffersson J., Grimsholm O., Jennbacken K., Sundqvist M., Wiseman J., Bidar A.W., Lindfors L., Synnergren J, & Valadi H. (2023). Lipid Nanoparticles Deliver the Therapeutic VEGFA mRNA In Vitro and In Vivo and Transform Extracellular Vesicles for Their Functional Extensions. Advanced Science, 10(12), 2206187.

+ Open protocol

+ Expand

Targeting HEY1 and RUNX2 in Chondrosarcoma

Check if the same lab product or an alternative is used in the 5 most similar protocols

shRNAs against human HEY1 and mouse Runx2 were lentivirally introduced into mouse mesenchymal chondrosarcoma cell lines. Knockdown efficiencies of each gene were confirmed by immunoblotting using the primary antibodies. The list of shRNAs is shown in Supplemental Table 8.
Mouse mesenchymal chondrosarcoma cells were treated with panobinostat in vitro. Cells were seeded into 96-well plates at a concentration of 5 × 10³ cells and were treated with drugs for 48 hours. Cell proliferation analysis was then performed using a XTT kit (Roche), and IC₅₀ was calculated. DNA synthesis analysis was performed using the Click-it EdU cell proliferation kit (Thermo Fisher Scientific). FACS analysis using the BD FACSLyric system (BD Bioscience) was performed for 48 hours after treatment with 10 nM drugs to detect EdU-positive fractions. For in vivo experiments, 5 × 5 mm tumor masses were transplanted subcutaneously into BALB/c nude mice, and the mice were treated with panobinostat or adriamycin (Selleckchem). panobinostat was intraperitoneally administered at 10 mg/kg 5 times continuously per week for 2 weeks, and adriamycin was intraperitoneally 3 mg/kg once a week for 2 weeks.

Tanaka M., Homme M., Teramura Y., Kumegawa K., Yamazaki Y., Yamashita K., Osato M., Maruyama R, & Nakamura T. (2023). HEY1-NCOA2 expression modulates chondrogenic differentiation and induces mesenchymal chondrosarcoma in mice. JCI Insight, 8(10), e160279.

+ Open protocol

+ Expand

Characterization of Cell Surface Markers

Check if the same lab product or an alternative is used in the 5 most similar protocols

To characterize the surface markers, the isolated cells were stained with fluorescence‐conjugated antibodies, which are listed in Table 2. Data acquisition and analysis were performed using the BD FACSLyric system (BD Biosciences) and FlowJo software (BD Biosciences).

Huang J., Lin Y., Wang L, & Chiang B. (2023). M2‐like macrophages polarized by Foxp3− Treg‐of‐B cells ameliorate imiquimod‐induced psoriasis. Journal of Cellular and Molecular Medicine, 27(11), 1477-1492.

+ Open protocol

+ Expand

CD132 Expression Evaluation in ED7R Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

ED7R cells, which do not express CD132, were cultured in RPMI 1640 (Life Technologies) with 10% FBS and 2 mM glutamine (R10 media) and plated at 2 × 10⁵ cells/well in a 12-well tissue culture plate. Purified LVVs and 8 μg/mL Polybrene (EMD Millipore) were added to each well for a final volume of 500 μL/well. The plates were centrifuged at 1,000 × g for 1 h and incubated at 37°C for 3 h before the addition of 1 mL of fresh R10 medium. After 72 h in culture, cells were harvested into a 96-well deep well plate. Cells were washed with phosphate-buffered saline (PBS) (Lonza) and incubated briefly on ice with rat immunoglobulin G (IgG) in PBS with 2% FBS. Biotin-labeled rat anti-human CD132 primary antibody (BD Biosciences, 10 μg/mL) in PBS with 2% FBS was added to the cells for 30 min on ice. The cells were washed with PBS, and 0.5 μg/mL of streptavidin phycoerythrin (PE)-conjugated secondary antibody (BD Biosciences) prepared in PBS with 2% FBS was added for 20 min on ice. Cells were washed with PBS, filtered through 30- to 40-μm 96-well AcroPrep Advance filter plates (Pall Corporation) and suspended in 150 μL of DAPI + PBS with 2% FBS for sorting by using the BD FACSLyric system. Cells that stably express CD132 were used as a positive control, and data were analyzed by linear regression of known titer values of an internal standard.

Powers A.D., Drury J.E., Hoehamer C.F., Lockey T.D, & Meagher M.M. (2020). Lentiviral Vector Production from a Stable Packaging Cell Line Using a Packed Bed Bioreactor. Molecular Therapy. Methods & Clinical Development, 19, 1-13.

+ Open protocol

+ Expand

Phenotypic Analysis of Immune Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

Fresh PBMCs from patients and healthy donors were washed and stained with the following antibodies at 4 °C for 15 min: anti-CD3-V450 (#560366, BD Biosciences), anti-CD4-V500 (#560769, BD Biosciences), anti-CD45RA-PE-Cy7 (#560675, BD Biosciences), anti-CXCR5-APC (#356907, BioLegend), and anti-PD-1-PE-Cy7 (#561272, BD Biosciences). For intracellular staining of anti-Foxp3-Alexa Fluor 488 (#560047, BD Biosciences), the cells were fixed and permeabilized with Transcription Factor Buffer at 4 °C for 30 min and were washed with Perm/Wash Buffer (BD Biosciences) before intracellular staining. Isotype-matched control antibodies were used to monitor the background. The well-stained cells were subjected to flow cytometry using a BD FACSLyric system and were further analyzed using the FlowJo v10 software (TOMY Digital Biology).

Hao H., Nakayamada S., Ohkubo N., Yamagata K., Zhang M., Shan Y., Iwata S., Zhang T, & Tanaka Y. (2021). Involvement of lncRNA IL21-AS1 in interleukin-2 and T follicular regulatory cell activation in systemic lupus erythematosus. Arthritis Research & Therapy, 23, 302.

+ Open protocol

+ Expand

Quantification and Characterization of Exosomal mRNA Delivery

Check if the same lab product or an alternative is used in the 5 most similar protocols

HTB-177 cells were treated with MC3-LNPs containing 100 µg of Cy5 mRNA (Trilink) as described above. Untreated cells were included as control. After 96 h, the total EVs were isolated by UC (pre-enrichment) and quantified. After pre-enrichment, the CD63 / CD9 positive EVs were isolated by using an affinity-based method and evaluated for the presence of Cy5 mRNA by FACS. In first place, exosome-Human CD63 isolation/detection reagent for cell culture medium (Thermo Fisher Scientific) was used to immobilize the CD63⁺ EVs to magnetic dynabeads according to manufacturer’s instructions. In the binding reaction, 20 µl of beads were incubated with 25 µg or 50 µg of total mc3-EVs or total untreated EVs. As negative control, 20 µl of beads were incubated with an equivalent volume of PBS (no EVs). After the CD63⁺ EVs immobilization, these EVs were stained with a mouse anti-human PE-CD9 antibody (BD Pharmingen™, cat. no. 555372) diluted 1:6 according to the manufacturer instructions. EVs were acquired on a BD FACSLyric system (BD Biosciences) and CD9 and Cy5 mRNA were detected and the data were analyzed using FlowJo software (TreeStar Inc.). The experiment was performed in biological duplicate. The gating strategy for beads by FACS analysis is represented in Supplementary Fig. 11a.

Maugeri M., Nawaz M., Papadimitriou A., Angerfors A., Camponeschi A., Na M., Hölttä M., Skantze P., Johansson S., Sundqvist M., Lindquist J., Kjellman T., Mårtensson I.L., Jin T., Sunnerhagen P., Östman S., Lindfors L, & Valadi H. (2019). Linkage between endosomal escape of LNP-mRNA and loading into EVs for transport to other cells. Nature Communications, 10, 4333.

+ 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?

Revolutionizing how scientists
search and build protocols!