Log In Sign up
The largest database of trusted experimental protocols

Sytox blue

Manufactured by Tree Star
Visit Supplier
Sourced in United States

Sytox Blue is a fluorescent dye used for nucleic acid staining. It binds to DNA and RNA, enabling the visualization and quantification of cellular nucleic acids in flow cytometry and fluorescence microscopy applications.

Automatically generated - may contain errors

Lab products found in correlation

Sytox blue, by Thermo Fisher Scientific (4 mentions) Collagenase, by Merck Group (3 mentions) F4 80 apc cy7, by BioLegend (3 mentions) Cd11c pe, by BD (3 mentions) Propidium iodide staining solution, by Merck Group (2 mentions) Cd31 pe cy7, by Thermo Fisher Scientific (2 mentions) F4 80 apc, by Thermo Fisher Scientific (2 mentions) Sca 1 fitc, by Thermo Fisher Scientific (2 mentions) Cd45 pe cy7, by Thermo Fisher Scientific (2 mentions) Cd206 alex647, by Bio-Rad (2 mentions) Facsaria cell sorter, by BD (2 mentions) Cd45 percp cy5, by BioLegend (1 mentions) 3 3 dihexyloxacarbocyanine iodide dioc6 3, by Thermo Fisher Scientific (1 mentions) Anti cd38 percp cy5.5, by BD (1 mentions) Anti cd5 apc, by BD (1 mentions) Anti cd27 pe cy7, by Thermo Fisher Scientific (1 mentions) Anti igd pe, by BD (1 mentions) Lsrii flow cytometer, by BD (1 mentions) Anti cd19 apc cy7, by BD (1 mentions)

4 protocols using sytox blue

1

Flow Cytometric Analysis of Adipose Tissue

Check if the same lab product or an alternative is used in the 5 most similar protocols
Stromal vascular fraction (SVF) was obtained from pgWAT by treatment with 2 mg/mL collagenase (Sigma) for 45 min at 37 °C. The isolated SVF was resuspended in cold Hank’s balanced salt solution (HBSS) with 2% fetal bovine serum (FBS). Cells were incubated with CD45-PE-Cy7 (eBiosciences), F4/80-APC-Cy7 (BioLegend), CD206-Alex647 (Serotec, Inc.) and CD11c-PE (BD Pharmingen) antibodies for 30 min in HBSS containing 2% FBS on ice and then washed and resuspended in solution with Sytox Blue (Thermo Scientific). Cells were analyzed on a BD FACSAria cell sorter after selection by forward scatter and side scatter, followed by exclusion of dead cells with Sytox Blue staining, and analyzed for cell-surface markers using FlowJo software (Tree Star). M1 or M2 macrophages were identified as F4/80-positive/CD11c-positive/CD206-negative or F4/80-positive/CD11c-negative/CD206-positive cells, respectively. The data are shown as the percentage of M1 and M2 macrophages. For sorting preadipocytes, endothelial cells and macrophages, cells were incubated with CD31-PE-Cy7 (eBiosciences), F4/80-APC (eBiosciences), and Sca-1 FITC (eBiosciences) antibodies for 30 min in HBSS containing 2% FBS on ice and then washed and resuspended in solution with Propidium Iodide Staining Solution (Sigma). Gating strategies are presented in Supplementary Figure 12 and Supplementary Figure 13.
Takahashi H., Alves C.R., Stanford K.I., Middelbeek R.J., Pasquale N.i.g.r.o., Ryan R.E., Xue R., Sakaguchi M., Lynes M.D., So K., Mul J.D., Lee M.Y., Balan E., Pan H., Dreyfuss J.M., Hirshman M.F., Azhar M., Hannukainen J.C., Nuutila P., Kalliokoski K.K., Nielsen S., Pedersen B.K., Kahn C.R., Tseng Y.H, & Goodyear L.J. (2019). TGF-β2 is an exercise-induced adipokine that regulates glucose and fatty acid metabolism. Nature metabolism, 1(2), 291-303.
+ Open protocol
+ Expand
2

Flow Cytometric Analysis of Adipose Tissue

Check if the same lab product or an alternative is used in the 5 most similar protocols
Stromal vascular fraction (SVF) was obtained from pgWAT by treatment with 2 mg/mL collagenase (Sigma) for 45 min at 37 °C. The isolated SVF was resuspended in cold Hank’s balanced salt solution (HBSS) with 2% fetal bovine serum (FBS). Cells were incubated with CD45-PE-Cy7 (eBiosciences), F4/80-APC-Cy7 (BioLegend), CD206-Alex647 (Serotec, Inc.) and CD11c-PE (BD Pharmingen) antibodies for 30 min in HBSS containing 2% FBS on ice and then washed and resuspended in solution with Sytox Blue (Thermo Scientific). Cells were analyzed on a BD FACSAria cell sorter after selection by forward scatter and side scatter, followed by exclusion of dead cells with Sytox Blue staining, and analyzed for cell-surface markers using FlowJo software (Tree Star). M1 or M2 macrophages were identified as F4/80-positive/CD11c-positive/CD206-negative or F4/80-positive/CD11c-negative/CD206-positive cells, respectively. The data are shown as the percentage of M1 and M2 macrophages. For sorting preadipocytes, endothelial cells and macrophages, cells were incubated with CD31-PE-Cy7 (eBiosciences), F4/80-APC (eBiosciences), and Sca-1 FITC (eBiosciences) antibodies for 30 min in HBSS containing 2% FBS on ice and then washed and resuspended in solution with Propidium Iodide Staining Solution (Sigma). Gating strategies are presented in Supplementary Figure 12 and Supplementary Figure 13.
Takahashi H., Alves C.R., Stanford K.I., Middelbeek R.J., Pasquale N.i.g.r.o., Ryan R.E., Xue R., Sakaguchi M., Lynes M.D., So K., Mul J.D., Lee M.Y., Balan E., Pan H., Dreyfuss J.M., Hirshman M.F., Azhar M., Hannukainen J.C., Nuutila P., Kalliokoski K.K., Nielsen S., Pedersen B.K., Kahn C.R., Tseng Y.H, & Goodyear L.J. (2019). TGF-β2 is an exercise-induced adipokine that regulates glucose and fatty acid metabolism. Nature metabolism, 1(2), 291-303.
+ Open protocol
+ Expand
3

Isolation and Characterization of M1 Macrophages

Check if the same lab product or an alternative is used in the 5 most similar protocols
SVF was obtained from iWAT and BAT by treatment with 2 mg/ml collagenase (Sigma) for 45 min at 37 °C. The isolated SVF was resuspended in cold Hank’s balanced salt solution (HBSS) with 2% fetal bovine serum (FBS). Cells were incubated with CD45-PerCP-Cy5.5, Clone: 30F-11 (BioLegend, 103131, 1:100), F4/80-APC-Cy7, Clone: BM8 (BioLegend, BL123117, 1:100), CD206-Alex647, Clone: MR5D3 (Bio Rad, MCA2235A647T, 1:50) and CD11c-PE, Clone: HL3 (BD Pharmingen, 553802, 1:100) antibodies for 30 min in HBSS containing 2% FBS on ice and then washed and resuspended in solution with Sytox Blue (Thermo Scientific, S34857, 1:1000). Cells were analyzed on a SH8005 (SONY) cell sorter after selection by forward scatter and side scatter, followed by exclusion of dead cells with Sytox Blue staining, and analyzed for cell-surface markers in FlowJo software (Tree Star). M1 macrophages were identified as F4/80+/CD11c+/CD206− cells. The data are shown as the percentage of total and M1 macrophages.
Sakaguchi M., Okagawa S., Okubo Y., Otsuka Y., Fukuda K., Igata M., Kondo T., Sato Y., Yoshizawa T., Fukuda T., Yamagata K., Cai W., Tseng Y.H., Sakaguchi N., Kahn C.R, & Araki E. (2022). Phosphatase protector alpha4 (α4) is involved in adipocyte maintenance and mitochondrial homeostasis through regulation of insulin signaling. Nature Communications, 13, 6092.
+ Open protocol
+ Expand
4

PBMC Viability Assessment Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols
Peripheral blood mononuclear cells (PBMCs) were purified by standard density gradient centrifugation using Ficoll-Hypaque (Atom Reactiva, Barcelona, Spain) and used immediately for ex-vivo SCD assays. B-cell death was evaluated by culturing PBMC (1 × 10 6 cells/ml) in Roswell Park Memorial Institute 1640 medium (RPMI-1640) supplemented with 10% of FBS (R10 medium; ThermoFisher Scientific) for 24 h. Then, PBMCs were harvested and incubated with 40 nmol/l of the potentiometric mitochondrial probe 3,3'-dihexyloxacarbocyanine Iodide (DIOC 6 (3)) (ThermoFisher Scientific) in R10 for 1 h at 37°C/5% CO 2 . After that, cells were stained with anti-CD19 APC/Cy7, anti-IgD PE, anti-CD38 PerCP-Cy5.5, anti-CD5 APC (BD Biosciences), and anti-CD27 PE/Cy7 (eBiosciences). Finally, 0.3 μm of Sytox Blue (ThermoFisher Scientific) was added and incubated for 10 min. Cells were acquired in a LSR-II flow cytometer (BD Biosciences). Live cells were identified as DIOC 6 (3) bright/Sytox Blue negative using FlowJo software (Tree Star Inc., Ashland, Oregon, USA).
Carrillo J., Negredo E., Puig J., Molinos-Albert L.M., Rodríguez de la Concepción M.L., Curriu M., Massanella M., Navarro J., Crespo M., Viñets E., Millá F., Clotet B, & Blanco J. (2018). Memory B cell dysregulation in HIV-1-infected individuals. AIDS (London, England), 32(2).
+ 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!