Anti sca1 percp cy5

Manufactured by Thermo Fisher Scientific

Sourced in United States

The Anti-sca1-PerCP/Cy5.5 is a flow cytometry antibody reagent that detects the Sca-1 (Stem Cell Antigen-1) protein marker. It is conjugated with the PerCP/Cy5.5 fluorescent label, allowing for its detection and quantification by flow cytometry.

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Close spelling variants of this product

PerCP-Cy5.5 (62 citations) Sca1-PerCP (2 citations) Rat anti-Sca1 PerCP-Cy5.5 (2 citations)

6 protocols using anti sca1 percp cy5

Comprehensive Immunophenotyping by Flow Cytometry

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Flow cytometry analysis was performed as previously described [28 (link), 30 (link)]. Samples were measured on a FACSCalibur or FACSCanto II (BD Bioscience, Franklin Lakes, USA) and analyzed with FlowJo (Tree Star, Ashland, USA). Dead cells were identified by using a Zombie Aqua™ Fixable Viability Kit (Biolegend, San Diego USA). Doublets were excluded in every analysis. Samples were stained with anti-sca1-APC (R&D Systems, Minneapolis, USA) anti-flk-1-PE (BD Bioscience, Franklin Lakes, USA), anti-B220-APC (BD Bioscience, Franklin Lakes, USA), anti-IgM-Per-CP/Cy5.5 (BD Bioscience, Franklin Lakes, USA), anti-CD11b-APC/Cy7 (BD Bioscience, Franklin Lakes, USA), anti-CD5-PE (BD Bioscience, Franklin Lakes, USA), anti-sca1-FITC (BD Bioscience, Franklin Lakes, USA), anti-flk1-PE-Cy7 (BD Bioscience, Franklin Lakes, USA), anti-CD3-FITC (BD Bioscience, Franklin Lakes, USA), anti-CD4-PE/Cy7 (eBioscience, Waltham, USA), anti-CD8-APC (Biolegend, San Diego, USA), anti-sca1-PerCP/Cy5.5 (eBioscience, Waltham, USA), anti-CD11b-PE (BD Bioscience, Franklin Lakes, USA), anti-CD115-APC (Biolegend, San Diego USA), anti-Gr1-FITC (BD Bioscience, Franklin Lakes, USA) and anti-CD45 FITC (BD Bioscience, Franklin Lakes, USA).

Steffen E., Mayer von Wittgenstein W.B., Hennig M., Niepmann S.T., Zietzer A., Werner N., Rassaf T., Nickenig G., Wassmann S., Zimmer S, & Steinmetz M. (2020). Murine sca1/flk1-positive cells are not endothelial progenitor cells, but B2 lymphocytes. Basic Research in Cardiology, 115(2), 18.

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Immunophenotypic Analysis of MSCs

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The antibodies used to analyse MSCs included anti-CD45-APC (Clone 30-F11, 1:100; BioLegend, USA), anti-Ter119-APC (Clone TER-119, 1:100; BioLegend, USA), anti-CD31-PE-cy7 (Clone MEC13.3, 1:100; BioLegend, USA), anti-PDGFRα-biotin (Clone APA5, 1:200; eBioscience, USA), PE anti-mouse CD51 antibody (Clone RMV-7, 1:100; eBioscience, USA), anti-LepR-biotin antibody (BAF497,1:100; R&D Systems, USA), and Anti-Sca1-Percp-cy5.5 (eBioscience, USA).

Song J., Li J., Yang F., Ning G., Zhen L., Wu L., Zheng Y., Zhang Q., Lin D., Xie C, & Peng L. (2019). Nicotinamide mononucleotide promotes osteogenesis and reduces adipogenesis by regulating mesenchymal stromal cells via the SIRT1 pathway in aged bone marrow. Cell Death & Disease, 10(5), 336.

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Isolation of Fetal Bone Marrow Cells

Cited 1 time

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The bone marrow of untreated and PNA/DNA NP treated Hbb^th-4/Hbb⁺ mice were collected 3 days post NP delivery on E18.5 as previously reported^{36 (link)}. Freshly isolated fetal bone marrow cells were suspended in ice-cold DMEM + (Dulbecco’s modified Eagle’s medium, 10 mM HEPES, 2% fetal bovine serum [FBS]) at 10⁷ cells ml⁻¹ and stained for 15 min on ice with anti-cKit-PE (eBioscience, Cat #12-1171-82), anti-Sca1-(PerCP)-Cy5.5 (eBioscience, Cat #45-5981-82), and FITC-conjugated lineage marker antibodies [CD4 (eBioscience, Cat #11-0041-82), CD8 (eBioscience, Cat #11-0081-82), Ter119 (eBioscience, Cat #11-5921-82), Gr-1 (eBioscience, Cat #11-5931-82), and CD45 (eBioscience, Cat #11-0452-82)] (Thermo Scientific; Rockford, IL). All antibodies were used at a dilution of 1:100. Samples were then washed with 10 × volume of HBSS + (Hank’s balanced salt solution, 10 mM HEPES, 2% FBS) and centrifuged at 0.4 × g for 8 min at 4 °C. Cell pellets were resuspended in DMEM + and samples were immediately sorted by flow cytometry (BD FACSAria).

Ricciardi A.S., Bahal R., Farrelly J.S., Quijano E., Bianchi A.H., Luks V.L., Putman R., López-Giráldez F., Coşkun S., Song E., Liu Y., Hsieh W.C., Ly D.H., Stitelman D.H., Glazer P.M, & Saltzman W.M. (2018). In utero nanoparticle delivery for site-specific genome editing. Nature Communications, 9, 2481.

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Comprehensive Immune Phenotyping for Hematopoietic Stem and Progenitor Cells

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The immune surface phenotype was assessed for myeloid, lymphoid, mast cell, and multipotent HSPC marker: Anti-CD3

ε

/APC-Cy7 (BioLegend Catalogue no.100330), anti-B220-APC/Cy7 (BioLegend Catalogue No. 103224), anti-Ter119/APC-Cy7 (BioLegend Catalogue No. 103224), anti-CD11b/AF700 (BioLegend Catalogue No. 103224), anti-Gr1 (BioLegend Catalogue No. 108422), anti-Fc

ε

α

/FITC (eBioscience Catalogue No. 11-5898-85) or anti-Fc

ε

α

/PE (eBioscience, Catalogue No. 12-5898-83), anti-cKit/APC-Cy7 (BioLegend, Catalogue No. 105826), and anti-Sca-1/PerCP-Cy5.5 (eBioscience, Catalogue No. 45-5981-82). Samples were finally stained within 0.2

μ

g/mL DAPI (Sigma-Aldrich, Catalogue No. D9542) for dead cell exclusion. Flow cytometry was performed with the Cytoflex S (Beckman Coulter, Brea, CA, USA) cytometer, and data were analyzed with the CytExpert (Beckman Coulter) and FlowJo, Version 10.2 (Tree Star, Ashland, OR, USA) software.

Fleischauer J., Bastone A.L., Selich A., John-Neek P., Weisskoeppel L., Schaudien D., Schambach A, & Rothe M. (2023). TGFβ Inhibitor A83-01 Enhances Murine HSPC Expansion for Gene Therapy. Cells, 12(15), 1978.

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Isolation and Characterization of Epidermal Stem Cells

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Back skin was excised and adipose tissue was removed. The skin was placed on a layer of trypsin solution (0.25%, Life Technologies Germany) with the epidermal side facing upwards. After incubation for 2 hours, the epidermis was separated from the dermis using blunt forceps. The epidermis was minced with scalpels and digested in 10 ml trypsin solution (0.25%, Life Technologies) for another hour. The digest was stopped by adding an equal volume of medium (DMEM (2/3) + HAM`s F12 (1/3)) supplemented with 10% calcium free FBS and the cells were strained through a 40 μm nylon mesh. Cells were then centrifuged for 8 min at 500 g at RT and the pellet was resuspended in 2 ml FACS buffer. Epidermal stem cell populations were stained according to Jensen et al. [35 (link)]. For flow cytometry, cells were stained using the following antibodies: anti-CD45-FITC (1:400, eBioscience), anti-CD49f-PE (1:200, eBioscience), anti-CD34-eF660 (1:50, eBioscience), anti-Sca1-PerCP-Cy5.5 (1:200, eBioscience), anti-CD117-APC-Cy7 (1:800, Biolegend). Cell sorting and analysis was performed on an ARIA III cell sorter (BD) and data were recorded using DIVA software (BD) and analyzed using FlowJo (FlowJo, LLC).

Hiller B., Hoppe A., Haase C., Hiller C., Schubert N., Müller W., Reijns M.A., Jackson A.P., Kunkel T.A., Wenzel J., Behrendt R, & Roers A. (2018). Ribonucleotide excision repair is essential to prevent squamous cell carcinoma of the skin.. Cancer research, 78(20), 5917-5926.

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Flow Cytometry Antibody Panel

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Antibodies for flow cytometry were purchased from eBioscience company as follows: anti-CD45-FITC (11-0451); anti-F4/80-APC (17-4801); anti-MHCII-PE-Cy7 (25-5321); anti-CD11B-PE (12-0112); anti-Ly6G-PercpCy5.5 (45-5931); anti-EpCAM-APC (17-5791); anti-CD24-PE (12-0241); anti-Sca-1-Percp-Cy5.5 (45-5981);anti-CD4-APC (17-0042); anti-Tcrb-PercpCy5.5 (45-5961); anti-IL-17A-PE (12-7178). Cell suspensions were prepared by sieving and gentle pipetting. Cells were washed in ice-cold FACS buffer (2% FCS, 2 mM EDTA in PBS), then incubated with each antibody for 30 min and washed twice with FACS buffer. Data were acquired on a Beckman Gallios flow cytometer and cells were sorted by Beckman Moflo Astrios. Flowjo software was used for data analysis.

Yang D., Chen X., Wang J., Lou Q., Lou Y., Li L., Wang H., Chen J., Wu M., Song X, & Qian Y. (2019). Dysregulated Lung Commensal Bacteria Drive Interleukin-17B Production to Promote Pulmonary Fibrosis through Their Outer Membrane Vesicles. Immunity, 50(3).

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