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Streptavidin pe cy7

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Streptavidin PE-Cy7 is a fluorescent dye-conjugated protein that binds to biotin with high affinity. It is commonly used in flow cytometry and other immunoassays to detect and quantify biotinylated molecules.

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Lab products found in correlation

Lsrfortessa, by BD (6 mentions) Collagenase dispase, by Roche (3 mentions) Dnase 1, by Roche (3 mentions) Anti cd45 clone 30 f11, by Thermo Fisher Scientific (3 mentions) Anti epcam1 clone g8, by Thermo Fisher Scientific (3 mentions) Anti ly51 clone 6c3, by BioLegend (3 mentions) Cd104 clone 346 11a, by BioLegend (3 mentions) Facsaria fusion 1 cell sorter, by BD (3 mentions) Anti cd80 clone 16 10a1, by BioLegend (3 mentions) Anti mhc 2 clone m5 114, by Thermo Fisher Scientific (3 mentions) Biotinylated uea 1, by Vector Laboratories (3 mentions) Facsaria 2, by BD (2 mentions) Streptavidin pe, by Thermo Fisher Scientific (2 mentions) Live dead fixable near ir dead cell stain kit, by Thermo Fisher Scientific (2 mentions) Tcrvα2 b20, by Thermo Fisher Scientific (2 mentions) Facsaria, by BD (2 mentions) Lsr fortessa cytometer, by BD (2 mentions) Sytox blue, by Thermo Fisher Scientific (2 mentions) Ls column, by Miltenyi Biotec (2 mentions) Anti cd45 beads, by Miltenyi Biotec (2 mentions)

Spelling variants of this product

Streptavidin-PE (135 citations) PE-Cy7-conjugated-Streptavidin (4 citations)

34 protocols using streptavidin pe cy7

1

Multicolor Flow Cytometry of Hematopoietic Cells

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Antibodies against CD34-FITC (cat. no. 11-0341-82), Sca1-PE (cat. no. 12-5981-82), CD117-APC (cat. no. 17-1171-82), CD11b-biotin (cat. no. 13-0112-82), Gr1-biotin (cat. no. 13-5931-82), B220-biotin (cat. no. 13-0452-82), TER119-biotin (cat. no. 13-5921-82), and streptavidin-PE-Cy7 (cat. no. 25-4317-82) were purchased from Thermo Fisher Scientific, Inc. Biotinylated antibodies were detected using streptavidin-PE-Cy7. Antibodies against CD19-FITC (cat. no. 115505), NKp46-PE (cat. no. 137603), and CD3e-AF488 (cat. no. 300319) were obtained from BioLegend, Inc.
Hattori K., Takagi H., Ogata Y., Yamada T., Horiba H., Fukata K., Sakaida T., Yashiro Y., Hasegawa S, & Tanaka H. (2022). Immunostimulatory effects of a subcritical water extract of Ganoderma. Biomedical Reports, 18(1), 1.
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2

Isolation and Identification of Cardiac Immune Cells

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Freshly excised hearts were minced on ice and digested in serum free Iscoves DMEM supplemented with 2mg/mL of Collagenase Type 2 (Worthington, Cat #LS004176) and 0.02mg/mL of Deoxyribonuclease I from bovine pancreas (Sigma, Cat #DN25-100MG) at 37°C for 30 minutes. These cell suspensions were then neutralized with IDMEM containing 10% FBS and filtered through 0.45μm strainer before blocking with 2.4G2 (BD Pharmingen, Cat #553141) for 30 minutes on ice, and staining with Gr-1-APC (Clone RB6-8C5, eBioscience), CD11b-APC-Cy7 (M1/70, BD Pharmingen), Ly 6C-PerCP-Cy5.5 (HK1.4, eBioscience), Ly6G-FITC (1A8, eBioscience), MHC-II PE-Cy5 (M5/114.15.2, eBioscience), MerTK-PE (DS5MMER, eBIoscience), F4/80-PE-Cy5 (BM8, eBioscience) and CD64-Biotin (X54-5/7.1, Biolegend and used in conjunction with Streptavidin PE-Cy7, eBioscience) antibodies for 30 minutes on ice protected from light. Cells were washed with PBS prior to flow cytometry. The samples were then analyzed on BD LSR-II (UAB Flow Cytometry Core Facility). Neutrophils were identified as Gr-1+CD11b+Ly-6G+F4/80-MHC-II-. Cardiac macrophages were first gated on F4/80+CD11b+ cells and further gated for expression of MerTK and CD64. Flow cytometry data were analyzed using FlowJo software. Percent neutrophils and macrophages were determined as (Percent live gated cells from the heart) x (percent positive).
Shenoy A.T., Brissac T., Gilley R.P., Kumar N., Wang Y., Gonzalez-Juarbe N., Hinkle W.S., Daugherty S.C., Shetty A.C., Ott S., Tallon L.J., Deshane J., Tettelin H, & Orihuela C.J. (2017). Streptococcus pneumoniae in the heart subvert the host response through biofilm-mediated resident macrophage killing. PLoS Pathogens, 13(8), e1006582.
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3

Quantification of Lin⁻CD45⁻Vybr⁺ Cells

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To determine the amount of LinnegCD45negVybrpos (CH) cells, PBMCs derived from fractured and naive mice were collected in FACS buffer (PBS containing 2% heat-inactivated foetal bovine serum) and stained for 10 min at 4 °C with Lineage Cell Detection Cocktail-Biotin antibody (Lin), (Miltenyi Biotec, Bergish Gladbach, Germany). A Streptavidin PE-Cy7 (eBioscience, San Diego, CA, USA) was used for indirect staining to detect the biotinylated antibody cocktail. After washing, cells were stained with APC rat anti-mouse CD45 antibody (clone 30-F11) (BD Biosciences). A double staining with Vybrant DyeCycle Green Stain and Sytox AADvanced Dead Cell Stain (Molecular Probes, Milan, Italy) was used to discriminate debris, live and dead cells. A set of microsphere suspensions (2, 4, 6 μm) (Molecular Probes) was used as size references.
CH cells were sorted, with high purity mask, from naive and RFP-Tg mice. All experiments were performed on BD FACSAria II. Data were analyzed using BD FACSDiva software.
Lo Sicco C., Tasso R., Reverberi D., Cilli M., Pfeffer U, & Cancedda R. (2015). Identification of a New Cell Population Constitutively Circulating in Healthy Conditions and Endowed with a Homing Ability Toward Injured Sites. Scientific Reports, 5, 16574.
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4

Flow Cytometric Analysis of T Cell Surface Markers

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Analysis by flow cytometry of the cell surface marker profile was conducted on T cells exposed to ASC conditioned medium. A total of 3 × 105 cells were concentrated by centrifugation at 300g for 5 minutes, suspended in 50 μl PBS and labeled with the primary antibodies. The following primary antibodies were used: Anti-mouse CD3-biotin (eBiosciences, San Diego, CA, www.ebioscience.com), anti-mouse CD4-PE (eBiosciences), and anti-mouse CD8-PeCy5 (eBiosciences). The samples were incubated for 30 minutes at room temperature. After three washes with PBS, cells were incubated in secondary antibody streptavidin-PeCy7 (eBiosciences) for 15 minutes at room temperature. Additional analysis was performed with anti-mouse CD62L-APC/Cy7 (Biolegend, San Diego, CA, www.biolegend.com) and anti-mouse CD44-Alexa fluor 700 (Biolegend). The samples were then analyzed with Galios Flow Cytometer (Beckman Coulter, Brea, CA) running Kaluza software (Beckman Coulter). To assay cells by forward and side scatter of light, FACScan was standardized with microbeads (Dynosphere uniform microspheres; Bangs Laboratories Inc.; Thermo Fisher Scientific; Waltham, MA). At least 10,000 events were analyzed and compared with unstained cells.
Strong A.L., Bowles A.C., Wise R.M., Morand J.P., Dutreil M.F., Gimble J.M, & Bunnell B.A. (2016). Human Adipose Stromal/Stem Cells from Obese Donors Show Reduced Efficacy in Halting Disease Progression in the Experimental Autoimmune Encephalomyelitis Model of Multiple Sclerosis. Stem cells (Dayton, Ohio), 34(3), 614-626.
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5

Comprehensive Immune Cell Profiling

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Peritoneal lavage and splenocytes were obtained as described previously (6 (link)). The following fluorochrome-labeled antibodies were purchased from BD Biosciences (Heidelberg, Germany): αCD4 (L3T4 and RM4-5), αB220 (RA3-6B2), αCD11b (M1/70), αCD11c (HL3), αCD69 (H1.2F3), αCTLA-4 (UC10-4F10-11), and αLy6G (1A8). A biotinylated cocktail of antibodies against Armenian and Syrian hamster IgG was purchased from Biolegend (San Diego, Calif). Antibodies directed against CD19 (1D3), CD25 (PC61.5), and MHCII (M5/114.15.2) were acquired from eBioscience (Hatfield, UK). For intranuclear Foxp3 staining, we used αFoxp3 antibodies (3G3) and the Foxp3-Staining Buffer Set from Miltenyi Biotech (Bergisch Gladbach, Germany). Streptavidin-PE and Streptavidin-PE-Cy7 were obtained from eBioscience.
Schmoeckel K., Traffehn S., Eger C., Pötschke C, & Bröker B.M. (2015). Full Activation of CD4+ T Cells Early During Sepsis Requires Specific Antigen. Shock (Augusta, Ga.), 43(2), 192-200.
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6

Characterization of Reprogramming Cells

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Cells harvested at different time points of reprogramming were stained in FACS buffer for 30 min at 4 °C and washed with FACS buffer (1% FCS in PBS) prior to acquisition with LSR Fortessa (BD Biosciences) cytometer. The following antibodies from eBioscience were used: ICAM1-biotin (13-0541-82; Dilution: 1/100), CD44-APC (17-0441-82; Dilution 1/300), streptavidin-PE-Cy7 (25-4317-82; Dilution: 1/1500). Dead cells were excluded using LIVE/DEAD™ Fixable Near-IR Dead Cell Stain Kit (ThermoFisher Scientific, Dilution: 1/1500). Data were analyzed using Flowjo v10.
Kaemena D.F., Yoshihara M., Beniazza M., Ashmore J., Zhao S., Bertenstam M., Olariu V., Katayama S., Okita K., Tomlinson S.R., Yusa K, & Kaji K. (2023). B1 SINE-binding ZFP266 impedes mouse iPSC generation through suppression of chromatin opening mediated by reprogramming factors. Nature Communications, 14, 488.
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7

Multiparameter Flow Cytometry Analysis

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Flk1-bio, TIE2-bio, CD31-bio, c-KIT-APC, VE-CADHERIN-APC, CD41-PE and Streptavidin-PECy7 antibodies were purchased from eBioscience. The stained cells were sorted on ARIA or Influx flow cytometers or analysed on a FACS LSRII (Becton Dickinson). The staining data were analysed using the FlowJo software (TreeStar).
Stefanska M., Batta K., Patel R., Florkowska M., Kouskoff V, & Lacaud G. (2017). Primitive erythrocytes are generated from hemogenic endothelial cells. Scientific Reports, 7, 6401.
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8

Multiparameter Flow Cytometry Staining

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For cell surface molecules, mAb 24G2 was used to block FcRs and cells were stained using α-F4/80-biotin, α-Ly6C-FITC, α-Ly6G-APC/Cy7 (clone 1A8), α-MHCII-Alexa Fluor 700 from BioLegend (San Diego, CA) and α-CD11b-eFluor 450, α-CD11c-PE/Cy5, α-DEC205-APC, α-pan-NK CD49b-PE (clone DX5) and streptavidin-PE/Cy7 from eBioscience (San Diego, CA). All cell events were acquired on an LSRII flow cytometer (BD Biosciences, San Jose, CA) and analyzed using FlowJo (Tree Star, Ashland, OR).
Weinkopff T., de Oliveira C.I., de Carvalho A.M., Hauyon-La Torre Y., Muniz A.C., Miranda J.C., Barral A, & Tacchini-Cottier F. (2014). Repeated Exposure to Lutzomyia intermedia Sand Fly Saliva Induces Local Expression of Interferon-Inducible Genes Both at the Site of Injection in Mice and in Human Blood. PLoS Neglected Tropical Diseases, 8(1), e2627.
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9

Multiparametric T Cell Analysis

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Thymus and spleen were mechanically teased with glass slides to acquire single cell suspensions and cells were stained with antibodies to the following: CD45.1 (A20, eBioscience), CD45.2 (104, eBioscience), CD4 (RM4‐5, Biolegend), CD8 (53‐6.7, Biolegend), CD25 (PC61.5, Biolegend), CD44 (IM7, eBioscience), Qa2 (695H1‐9‐9, Biolegend), TCRβ (H57‐597, eBioscience), CD69 (H1.2F3, eBioscience), and H‐2Kb (AF6‐88.5, Biolegend). Reagents were conjugated to Pacific Blue, Brilliant Violet (BV) 421, BV510, BV711, PE, PE‐Cy7, PerCP–eFluor 710, allophycocyanin– eFluor 780 and Alexa Fluor 700. Streptavidin PE‐Cy7 (eBioscience) was used to detect biotinylated antibodies. A Foxp3 fixation kit (eBioscience) was used in conjunction with anti‐Foxp3 (FJK‐16s, eBioscience) or the BD Cytofix/Cytoperm Kit (BD Biosciences) to preserve the GFP signal. Data were acquired using a BD LSR Fortessa and FACSDiva 2.6 software and analysed using FloJo software (Tree Star).
Cowan J.E., Baik S., McCarthy N.I., Parnell S.M., White A.J., Jenkinson W.E, & Anderson G. (2018). Aire controls the recirculation of murine Foxp3+ regulatory T‐cells back to the thymus. European Journal of Immunology, 48(5), 844-854.
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10

Multicolor Flow Cytometry Antibodies

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Most fluorochrome-conjugated antibodies, unless otherwise indicated, were purchased from Biolegend (San Diego, CA) or eBioscience (San Diego, CA), including: CD3 (17A2), CD8 (53–6.7), CD11b (M1/70), CD11c (N418), CD16 and CD32/Fc block (93), CD19 (1D3), CD45R/B220 (RA3-6B2), CD115/CSF-1R (AFS98), Gr-1 (RB6-8C5), Ly-6C (HK1.4), MHC-II (M5/114.15.2), NK1.1 (PK136), Ter 119 (TER-119), TCRVα2 (B20.1), and TCRVβ5.1 (MR9-4). Streptavidin–PE-Cy7, streptavidin–PerCP–Cy5.5, and 7-amino-actinomycin D (7-AAD) viability staining solutions were obtained from eBioscience, and streptavidin-APC-Cy7 was obtained from Biolegend. ExtrAvidin−TRITC was obtained from Sigma-Aldrich (St. Louis, MO).
Yang Y.W, & Luo W.H. (2017). Recruitment of bone marrow CD11b+Gr-1+ cells by polymeric nanoparticles for antigen cross-presentation. Scientific Reports, 7, 44691.
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