Pe cy7 conjugated streptavidin

Manufactured by Thermo Fisher Scientific

PE-Cy7-conjugated-Streptavidin is a fluorescently-labeled streptavidin reagent. Streptavidin is a tetrameric protein that binds to biotin with high affinity. The PE-Cy7 fluorescent label allows for detection and visualization of biotin-labeled targets.

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Close spelling variants of this product

Streptavidin-PE (135 citations) PE-conjugated streptavidin (40 citations) Streptavidin PE-Cy7 (34 citations)

4 protocols using pe cy7 conjugated streptavidin

Intracellular Cytokine Profiling of T Cell Subsets

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Mononuclear cells were incubated with or without 50 ng/ml phorbol
myristate acetate (PMA) (Sigma) and 500 ng/ml ionomycin (Sigma) in the presence
of GolgiStop (BD) in complete T cell media at 37°C for 5 h.
Intracellular cytokine staining was performed according to the
manufacturer’s protocol. Cells were stained with Pacific Blue-conjugated
anti-CD4 (RM-5), PerCP-Cy5.5-conjugated anti-CD8a (53-6.7), APC-Cy7-conjugated
anti-TCRβ (H57-597), FITC-conjugated anti-CD62L (MEL-14), APC-conjugated
anti-CD44 (IM7), PE-conjugated-CD25 (PC61.5), PerCP-Cy5.5-conjugated-CD19
(eBio1D3), APC-conjugated anti-CD45.1 (A20), FITC-conjugated anti-CD45.2 (104),
Pacific Blue-conjugated anti-CD11c (N418), FITC conjugated-anti-CD11b (M1/70),
PerCP-Cy5.5-conjugated-anti-CD103 (2E7) (eBioscience), Biotin-conjugated
Vβ14 (14-2) (BD phamigen) and PE-Cy7-conjugated-Streptavidin (Thermo
Fisher Scientific). Cells were further stained intracellularly with
APC-conjugated anti-RORγ (B2D) (eBioscience) and PE-Cy7-conjugated
anti-IL-17a (eBio17B7) (eBioscience) using Foxp3 staining/permeabilization
buffer (eBioscience). Flow cytometric analysis was performed on an LSRII (BD
Biosciences). All data were re-analyzed using FlowJo (Tree Star).

Kim S., Kim H., Yim Y.S., Ha S., Atarashi K., Tan T.G., Longman R.S., Honda K., Littman D.R., Choi G.B, & Huh J.R. (2017). Maternal gut bacteria promote neurodevelopmental abnormalities in mouse offspring. Nature, 549(7673), 528-532.

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Intracellular Cytokine Profiling of T Cell Subsets

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Isolation of Muscle and Tumor Cell Subsets

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Phenotypically distinct muscle and tumor cell subsets were sorted according to protocols that were previously established to isolate functionally discrete subsets of myofiber-associated cells [10 (link), 11 (link), 1 (link)]. In brief, cells were suspended in HBSS supplemented with 2% FBS. Antibody staining was performed for 20 minutes on ice. The following primary and secondary antibodies were used: APC-CY7-conjugated anti-mouse CD11b (1 in 200, BD Pharmingen, 557657), APC-CY7-conjugated anti-mouse CD45 (1 in 200, eBioscience, 557659), APC-CY7-conjugated anti-mouse TER119 (1 in 200, BioLegend, 116223), APC-conjugated anti-mouse Sca1 (1 in 200, eBioscience, 17-5981-82), PE-conjugated anti-mouse/ rat CD29 (1 in 400, BioLegend, 102207), biotin rat anti-mouse CD184 (1 in 100, BD Pharmingen, 551968), PECY7-conjugated Streptavidin (1 in 200, eBioscience, 25-4317-82). Antibody staining was performed for 20 minutes on ice. Prior to FACS sorting, cells were suspended in 1µg/ml propidium iodide and 10µM calcein blue (Invitrogen) to identify viable cells (Pi⁻Ca⁺). Cells were sorted twice to maximize purity.

Hettmer S., Lin M.M., Tchessalova D., Tortorici S.J., Castiglioni A., Desai T., Mao J., McMahon A.P, & Wagers A.J. (2015). Hedgehog-driven myogenic tumors recapitulate skeletal muscle cellular heterogeneity. Experimental cell research, 340(1), 43-52.

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Single Cell Flow Cytometry with Lipid Peroxidation

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Single cell suspensions at a concentration of 10⁶ cells/ml were incubated with 1 μM C11-BODIPY 581/591 (Thermo Fisher Scientific, Cat# D3861) for 30 min at 37 °C and 5% CO₂. Cells were then centrifuged and resuspended in staining buffer (2% fetal bovine serum in PBS) at a concentration of 10⁷ cells/ml. Two million cells (0.2 ml) were stained in quadruplicate for each staining condition. Cells were seeded in a 96-well plate, centrifuged, and resuspended in Fc Block in staining buffer for 10 min at 4 °C. The cells were then incubated for 30 min at 4 °C with appropriate antibodies diluted in staining buffer to achieve the final concentrations stated above, as well as LIVE/DEAD Fixable Violet Dead Cell Stain (Invitrogen, Cat# L34955) following manufacturer's recommendation. Cells were washed twice with staining buffer and stained for 30 min at 4 °C with PE/Cy7 Conjugated Streptavidin (eBiosciences, Cat# 25-4317-82) in staining buffer (1:1000). Cells were then washed twice with staining buffer, fixed in Fixation Buffer (BioLegend, Cat# 420801) for 15 min at room temperature, and then resuspended in staining buffer. Flow cytometry was subsequently performed using a BD LSRFortessa cell analyzer, and the data were analyzed using FlowJo software (www.flowjo.com).

Nunes L.G., Ma C., Hoffmann F.W., Shay A.E., Pitts M.W, & Hoffmann P.R. (2024). Selenoprotein I is indispensable for ether lipid homeostasis and proper myelination. The Journal of Biological Chemistry, 300(5), 107259.

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