Streptavidin apc cy7

Manufactured by BioLegend

Sourced in United States

Streptavidin-APC-Cy7 is a fluorescent conjugate used in flow cytometry and other immunoassays. Streptavidin, a protein that binds biotin, is conjugated to the tandem dye APC-Cy7, which emits fluorescence when excited by a laser. The product can be used to detect and quantify biotinylated molecules.

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Lab products found in correlation

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Spelling variants of this product

APC-Cy7 (106 citations) Streptavidin-APC (85 citations) APC-Cy7-conjugated streptavidin (6 citations) APC-Cy7- or BV421-labeled streptavidin-antibody (2 citations)

37 protocols using streptavidin apc cy7

Comprehensive Immunophenotyping Panel

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Alexa Fluor 488 NK1.1 (PK136), Biotin-CD4 (GK1.5), Alexa Fluor 488-CD4 (GK1.5), APC-eFluor 780-CD4 (GK1.5), FITC-CD3ε (17A2), Alexa Fluor 647-CD3ε (17A2), Alexa Fluor 488-CD206, Brilliant Violet 421-CD3ε (17A2), FITC-F4/80, Alexa Fluor 488-CD3ε (145-2C11), Alexa Fluor 647 CD49b (DX5), Pacific Blue-CD49b (DX5), PE-NK1.1 (PK136), Brilliant Violet 421 NK1.1 (PK136), PE/Cy5-CD4 (GK1.5), APC-TCRγδ (GL3), Biotin-IFN-γRα (2E2), Brilliant Violet 421-CD25 (PC61), PE-CD25 (PC61), Biotin-CD122 (5H4), PE/Cy7-IFN-γ (XMG1.2), PE-GM-CSF (MP1-22E9), Pacific Blue-TNF-α (MP6-XT22), PercCP/Cy5.5-CD69, Biotin-BrdU (Bu20a), FITC-Ki67 (16A8), Alexa Fluor 647-streptavidin, APC-Cy7-Streptavidin, PE-Cy7-Streptavidin, Brilliant violet 421-CD62L (MEL-14) and rabbit polyclonal anti-Asialo-GM1 (aGM) antibodies were purchased from Biolegend (San Diego, CA). APC-RORγt (AFKJS-9), APC-T-bet (4B10) and PE/Cy7-T-bet (4B10) were procured from eBioscience (San Diego, CA). Anti-NK1.1 (PK136), anti-F4/80 (Cl:A3–1), anti-Gr1 (RB6-8C5) and isotype control antibodies for in vivo use were procured from BioXcell (West Lebanon, NH). Alexa Fluor 647-labeled CD1d tetramer (mCD1d, PBS-57) and unloaded control tetramer were received from NIH Tetramer Core Facility (Atlanta, GA). 5-Bromo-2′-deoxyuridine (BrdU) was purchased from Sigma-Aldrich.

Paul S., Chhatar S., Mishra A, & Lal G. (2019). Natural killer T cell activation increases iNOS+CD206- M1 macrophage and controls the growth of solid tumor. Journal for Immunotherapy of Cancer, 7, 208.

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Isolation of Skeletal Stem Cells

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All limbs were collected from 8-week-old mice that had been subjected to tamoxifen and all muscles and ligaments removed. Cleaned bones (i.e., whole bone preparation) were gently disrupted using a mortar and pestle, minced, and digested in 2.5 mg/mL collagenase I (#CLS-1, Worthington). Collected cells were subjected to RBC lysis and blocked with 3% BSA, 2% FCS in PBS. The following antibodies were used: anti-CD45 (#103131, BioLegend, 1:80), anti-Ter119 (#116227, BioLegend, 1:80), anti-CD31 (#562861, BD Pharmingen, 1:80), anti-CD200 (#ab33735, Abcam, 1:10), APC/Cy7 Streptavidin (#405208, BioLegend, 1:300) and DAPI (#D9542, Sigma-Aldrich). Cells were sorted on FACSFusion or LSR Fortessa (BD Biosciences) based on forward and side scatter plots, single cells, live cells (DAPI negative), trilineage expression (CD45^-Ter119^-CD31^-) and lineage reporter positive fluorescence.

Ng J.Q., Jafarov T.H., Little C.B., Wang T., Ali A.M., Ma Y., Radford G.A., Vrbanac L., Ichinose M., Whittle S., Hunter D.J., Lannagan T.R., Suzuki N., Goyne J.M., Kobayashi H., Wang T.C., Haynes D.R., Menicanin D., Gronthos S., Worthley D.L., Woods S.L, & Mukherjee S. (2023). Loss of Grem1-lineage chondrogenic progenitor cells causes osteoarthritis. Nature Communications, 14, 6909.

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Multiparameter Flow Cytometry Staining

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Staining was performed with hybridoma supernatant 2.4G2 Fc block (ATCC HB197) in FACS staining buffer at 4°C. The following antibodies were purchased from Biolegend: biotin-anti-CD11c (N418), Brilliant Violet 650 anti-CD11c (N418), PE-anti-CD11b (M1/70), PE-anti-CD3 (17A2), PE-anti-CD8 (53–6.7), PE-anti-Gr-1 (RB6-8C5), PE-anti-CD19 (eBio1D3), PE-anti-TER119 (TER-119), PE-anti-Thy1.1 (HIS51), PE-anti-NK1.1 (PK136), APC-anti-B220 (RA3-6B2), FITC anti-TLR9 (M9.D6), Brilliant Violet 421 anti-B220 (RA3-6B2), PerCP-eFluor710-anti-Siglec-H (eBio440c), PerCP-eFluor710-anti-c-Kit (2B8), APC-anti-M-CSFR (AFS98), PE/Cy7-anti-IL-7Rα (A7R34), FITC-anti-Sca-1 (D7). PE-anti-MHCII (NIMR-4) and FITC-anti-MHCII (NIMR-4) were purchased from eBioscience, and FITC-anti-IFNα (RMMA-1) was purchased from PBL Assay Science. APC-Cy7-Streptavidin was used as the secondary antibody (Biolegend). Flow data were analyzed with FlowJo (FLOWJO LLC) software.

Chen Y.L., Chang S., Chen T.T, & Lee C.K. (2015). Efficient Generation of Plasmacytoid Dendritic Cell from Common Lymphoid Progenitors by Flt3 Ligand. PLoS ONE, 10(8), e0135217.

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Surface Marker Analysis of Immune Cells

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For surface markers analysis, live cells were re-suspended in 1 × PBS and stained with anti-mouse CD45 (30-F11, Biolegend), CD11b (M1/70, R&D Systems), F4/80 (BM8, Biolegend), CD86 (GL-1, Biolegend), MHC-II (M5/114.15.2, Biolegend), CD117 (2B8, Biolegend), CD115 (AFS98, Biolegend), CD16/32 (93, Biolegend), CD34 (HM34, Biolegend), Sca1 (D7, Biolegend), SIRPα (P84, biolegend), Lin (Stem Cell), and APC-Cy7 Streptavidin (Biolegend) at 4°C for 30 min. The concentration at each antibody was used as the product protocol recommended. For intracellular cytokine staining, cells were fixed and permeabilized with Fixation and Permeabilization Kit (eBioscience) at room temperature for 40 min and labeled with anti-mouse CD206 (C068C2, Biolegend). Multicolor FACS analysis was performed on an LSR Fortessa Analyzer (BD Biosciences). All data analysis was performed using the flow cytometry analysis program FlowJo-V10.

Zhao C.C., Han Q.J., Ying H.Y., Gu X.X., Yang N., Li L.Y, & Zhang Q.Z. (2022). TNFSF15 facilitates differentiation and polarization of macrophages toward M1 phenotype to inhibit tumor growth. Oncoimmunology, 11(1), 2032918.

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Osteoclastogenesis Regulation by Key Factors

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Recombinant mouse M-CSF, GM-CSF, IL-4, and RANKL were purchased from BioLegend (San Diego, CA). AF488-phalloidin was purchased from Invitrogen (Carlsbad, CA). Antibodies for immunoblot assays were obtained against NFATc1 (Pierce, Rockford, IL); Oscar (R&D Systems, Minneapolis, MN); TRAP (BioLegend); Integrin β3; c-Fos; cleaved Caspase-3, -8, and -9; and cleaved PARP (Cell Signaling Technology, Mountain View, CA). The previously described anti-Ninj1 Ab_1-15^{18 (link)} was used for immunoblotting assays. For FACS analysis, Fc block, and PE-anti-mouse CD115, APC-anti-mouse CD117, and V450-anti-mouse CD11b antibodies were purchased from BD Biosciences (Bedford, MA), and biotin-anti-RANK antibody and APC/Cy7 streptavidin were obtained from BioLegend.

Bae S.J., Shin M.W., Son T., Lee H.S., Chae J.S., Jeon S., Oh G.T, & Kim K.W. (2019). Ninjurin1 positively regulates osteoclast development by enhancing the survival of prefusion osteoclasts. Experimental & Molecular Medicine, 51(1), 1-16.

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Multicolor Flow Cytometric Profiling of Immune Cells

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Single cell suspensions prepared from lungs, BLNs, spleens and bronchoalveolar lavage were stained with the indicated combinations of Alexa Fluor 780-conjugated anti-CD4 (RM4–5, eBioscience), anti-CD8-PEcy7 (53-6.7, BioLegend), biotinylated anti-ICOS (7E.17G9, BioLegend), biotinylated anti-CD43 (1B11, BioLegend) and APC/cy7streptavidin (BioLegend). For intracellular cytokine staining, cells were stimulated for 2 hours with phorbol 12-myristate 13-acetate (PMA) (0.1 µg/ml, Cat#p1585, Sigma) and ionomycin (1 µg/ml, Cat#I0634, Sigma) and with golgi stop (Brefeldin) (5 µl/ml, Cat#B7651, Sigma) for 2 hours, followed by permeabilization with NP-40 (igepal CA-630, Sigma) and staining with anti-IFNγ-APC (XMG1.2, BioLegend). Intracellular FoxP3 expression was determined using the FoxP3 staining buffer kit (FJK-16s, eBioscience) according to the manufacturer’s recommendations. For apoptosis assay, Annexin/7AAD staining was performed as described previously [35] (link). Flow cytometric analysis was performed using a FACSCanto or LSRFortessa (BD Biosciences) running with THE FACSDiva software (version 6.1.3) (BD Biosciences). Analysis was performed using FlowJo version 9.5.3 (Tree star, Inc.).

Sakthivel P., Gereke M., Breithaupt A., Fuchs D., Gigliotti L., Gruber A.D., Dianzani U, & Bruder D. (2014). Attenuation of Immune-Mediated Influenza Pneumonia by Targeting the Inducible Co-Stimulator (ICOS) Molecule on T Cells. PLoS ONE, 9(7), e100970.

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Multiparametric analysis of hematopoietic cells

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Single cell suspensions were prepared from bone marrow and spleen cells as previously described^{19 (link)}. APC-c-kit, PE-Cy7-Sca-1 and Pacblue-Ki67 antibodies were purchased from eBioscience. APC-Cy7-Streptavidin, FITC-CD45.1 and APC-CD45.2 antibodies were purchased from Biolegend. The bone marrow biotin lineage panel and 7AAD/annexin-V staining kit were purchased from BD Pharmingen. All flow samples were analyzed by Canto-II or Fortessa flow cytometer and sorted by Aria-II cell sorter (Becton Dickinson).
For phospho-Flow, bone marrow cells were stimulated with or without a cytokine cocktail, containing murine IL-3 (PeproTech) 10ng/mL, murine IL-6 (PeproTech) 10ng/mL, murine SCF (PeproTech) 100ng/mL, and murine GM-CSF (BioLegend) 200 ng/mL for 15 minutes. Cells were then fixed in 1.5% paraformaldehyde (Electron Microscopy Sciences) at 37 °C for 10 minutes and permeabilized on ice with Methanol for 15 minutes. Subsequently, cells were stained with antibodies to cell surface markers as well as to pERK1/2 (T202/Y204), pSTAT3 (Y705), pSTAT5 (Y694), pAKT (S473) or IgG isotope control for 30 minutes at room temperature, before being subjected to FACS analysis. All phospho-specific antibodies were purchased from eBiosciences and are PE-conjugated.

Cheng G., Liu F., Asai T., Lai F., Man N., Xu H., Chen S., Greenblatt S., Hamard P.J., Ando K., Martinez C., Tadi M., Wang L., Xu M., Yang F.C., Shiekhattar R, & Nimer S.D. (2016). Loss of p300 Accelerates MDS-associated Leukemogenesis. Leukemia, 31(6), 1382-1390.

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Immunophenotyping of Bone Marrow DCs

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Bone marrow-derived DCs were incubated on ice with fluorochrome- or biotin-conjugated CD11b, CD11c, and AF6-88.5 antibodies diluted in PBS with 2% FCS. APC/Cy7-streptavidin (Biolegend) was used as a secondary reagent and 7-AAD (5 μl/sample) to discriminate live and dead cells. BD Fortessa^TM and fluorescence-activated cell sorting (FACS) ARIA-II^TM machines were used for cell analysis and sorting, respectively.

Weimershaus M., Mauvais F.X., Evnouchidou I., Lawand M., Saveanu L, & van Endert P. (2020). IRAP Endosomes Control Phagosomal Maturation in Dendritic Cells. Frontiers in Cell and Developmental Biology, 8, 585713.

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Flow Cytometry Analysis of Reprogrammed Cells

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Cells from standard reprogramming, Colony Forming Unit (CFU), or LTC experiments were first harvested using trypLE express at specified days and washed with PBS supplemented with 5% FBS and 1mM EDTA. Flow cytometry analysis was performed on a 5-laser LSRII with Diva software (BD Biosciences) and analyzed with FCS Express 6 Flow Research Edition (Win64). Cells were stained with PE/CY7-hCD45 (2D1), FITC-hCD235a (GA-R2), APC-hCD41 (MReg30), BV421-hCD14 (M5E2), BV421-hCD34 (581), APC-hCD45 or FITC-hCD45 (2D1), PE-hEPCR (RCR-401), or APC-hCD49f (GoH3) (all Biolegend), as well as PE-hACE (BB9), FITC-hCD90 (5E10), PE-hCD49f (GoH3) (BD Biosciences), APC-hCD90 (5E10, Thermo Fisher), or hACE-Biotin (BB9, R&D Scientific Corporation) with APC-Cy7 Streptavidin (BioLegend). 4,6-diamidino-2-phenylindole (DAPI, 1ug/mL, Sigma) or Propidium Iodide (PI, 1ug/ml, R17755, Invitrogen) was added prior to analysis to exclude dead cells. Sorting for RNA sequencing, transplants, LTC, and CFU assays was performed with APC-CD49f alone, PE-CD49f alone or BV421-CD34 and PE-CD49f using DAPI or PI to exclude dead cells.

Daniel M.G., Sachs D., Bernitz J.M., Fstkchyan Y., Rapp K., Satija N., Law K., Patel F., Gomes A.M., Kim H.S., Pereira C.F., Chen B., Lemischka I.R, & Moore K.A. (2019). Induction of human hemogenesis in adult fibroblasts by defined factors and hematopoietic co-culture. FEBS letters, 593(23), 3266-3287.

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Comprehensive Thymocyte Immunophenotyping

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Anti-CD4–Horizon V450, anti-CD4–FITC, anti-CD8–FITC, anti-CD8–Alexa647, anti-CD24–PE, anti-CD25–Alexa700, anti-CD25–APC-Cy7, anti-CD44–Horizon V450, anti-CD44–PE-Cy7, anti-CD45.1–Pacific Blue, anti-CD45.2–PE-Cy5, anti-CD117–PE-Cy7, anti-Sca-1–Alexa700, anti-TCRβ–PE, anti-TCRβ–PE-Cy5, and anti-Ter119–FITC were purchased from BioLegend (San Diego, CA) or BD Biosciences (San Jose, CA). Lineage markers were identified using biotinylated anti-Ter119, anti-CD11b, anti-Gr-1, anti-CD3ε, and anti-B220 followed by streptavidin-APC-Cy7 (Biolegend). Cell labeling was performed in PBS containing 2 % FCS. To quantify the number of thymocytes and mast cells, CountBright™ beads (Life Technologies, Grand Island, NY) were added.
Flow cytometry studies were performed using a BD LSR II (BD Immunocytometry Systems, San Jose, CA). Data were analyzed using BD FACSDiva software (BD Biosciences, San Jose, CA). Thymocytes were gated on the live lymphocyte gate, and doublet discrimination was performed. Thymocytes were gated on the Ter119⁻CD45⁺ population.

Xiong J., Parker B.L, & Yankee T.M. (2014). The combined loss of Gads and CD127 reveals a novel function of Gads prior to TCRβ expression. Immunologic research, 60(1), 77-84.

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