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Legendscreen mouse pe kit

Manufactured by BioLegend
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The LEGENDScreen Mouse PE Kit is a laboratory equipment product that is used for flow cytometry analysis. It provides a standardized method for the simultaneous screening of multiple mouse cell surface markers.

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Lab products found in correlation

Facs lysing solution, by BD (1 mentions) Live dead fixable aqua dead cell stain kit, by Thermo Fisher Scientific (1 mentions) Cytofix cytoperm kit, by BD (1 mentions) Facscanto 2, by BD (1 mentions) Cd20 clone 2h7, by BioLegend (1 mentions) Cd81 clone 5a6, by BioLegend (1 mentions) Cd39 clone a1, by BioLegend (1 mentions) Cd138 clone mi15, by BioLegend (1 mentions) Il 6 clone mp5 20f3, by BioLegend (1 mentions) Flow count fluorosphere, by Beckman Coulter (1 mentions)

Close spelling variants of this product

LEGENDScreen kit LEGENDScreen

5 protocols using legendscreen mouse pe kit

1

Multicolor Flow Cytometry of Murine Splenocytes

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Single-cell leukocyte suspensions from spleens were generated by gentle dissection. The following mAbs were used: fluorescein isothiocyanate–, PE (phycoerythrin)–, PE-Cy5–, PE-Cy7–, PerCP-Cy5.5–, APC (allophycocyanin)–, APC-PECy7–, and BV421-conjugated mAbs to mouse CD4 (RM4-5), CD8 (53-6.7), CD11b (M1-70), CD19 (1D3), CD25 (MF-14), CD44 (IM7), IL-10 (MP5-20F3), and IL-10 (JES5-16E3), using LEGENDScreen Mouse PE Kit from BioLegend; CD1d (1B1), CD5 (53-7.3), CD21/CD35 (7G6), CD23 (B3B4), CD24 (M1/69), and CD45R/B220 (RA3-6B2) from BD Biosciences; and BAFF (121808) from R&D Systems. For two- to six-color immunofluorescence analysis, single-cell suspensions (106 cells) were stained at 4°C using predetermined optimal concentrations of mAb for 20 min. Blood erythrocytes were lysed after staining using FACS Lysing Solution (Becton Dickinson). For intracellular staining, cells were fixed and permeabilized with a Cytofix/Cytoperm kit (BD Biosciences). Dead cells were detected by using LIVE/DEAD Fixable Aqua Dead Cell Stain Kit (Invitrogen Molecular Probes) before cell surface staining. Cells with the forward and side light scatter properties of lymphocytes were analyzed using a BD FACSCanto II (BD Biosciences). Data were analyzed with FlowJo software (version 10.2; Tree Star).
Matsushita T., Kobayashi T., Mizumaki K., Kano M., Sawada T., Tennichi M., Okamura A., Hamaguchi Y., Iwakura Y., Hasegawa M., Fujimoto M, & Takehara K. (2018). BAFF inhibition attenuates fibrosis in scleroderma by modulating the regulatory and effector B cell balance. Science Advances, 4(7), eaas9944.
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2

Murine and Human Monoclonal Antibody Panel

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Murine specific monoclonal antibodies: CD3 (clone 17A2), CD11a (clone M17/4), CD18 (clone M18/2), CD16/CD32 (Fc Block, clone 2.4G2), CD19 (clone 6D5), CD39 (clone Duha59), CD44 (clone IM7), CD47 (clone miap301), CD49d (clone R1-2), CD54 (clone YN1/1.7.4), CD81 (clone Eat-2), CD98 (clone RL388), CD130 (clone 4H1B35), CD138 (clone 281-2), CD155 (clone TX56), CD172a (clone P84), CD205 (clone NLDC- 145), CD229 (clone Ly9ab3), CD267 (clone 8F10), CD274 (clone 10F.9G2), CD319 (clone 4G2), CD326 (clone G8.8), IgA (clone mA-6E1), IgM (clone RMM-1), LAG-3 (Clone eBioC9B7W), SCA-1 (clone D7) and human specific monoclonal antibodies: CD3 (clone UCHT1), CD14 (clone M5E2), CD19 (clone SJ25C1), CD20 (clone 2H7), CD27 (clone L128), CD39 (clone A1), CD81 (clone 5A6), CD130 (clone 2E1B02), CD138 (clone MI15), and CD326 (clone 9C4) were purchased from BioLegend, Miltenyi Biotec, BD Biosciences, Thermo Fisher or produced in house. LEGENDScreen™ Mouse PE Kit was purchased from BioLegend.
Dang V.D., Mohr E., Szelinski F., Le T.A., Ritter J., Hinnenthal T., Stefanski A.L., Schrezenmeier E., Ocvirk S., Hipfl C., Hardt S., Cheng Q., Hiepe F., Löhning M., Dörner T, & Lino A.C. (2022). CD39 and CD326 Are Bona Fide Markers of Murine and Human Plasma Cells and Identify a Bone Marrow Specific Plasma Cell Subpopulation in Lupus. Frontiers in Immunology, 13, 873217.
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3

Comprehensive Immunophenotyping of Murine Cells

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Monoclonal anti-mouse CD11b (clone M1/70), CX3CR1 (clone SA011F11), F4/80 (clone BM8), NK1.1 (clone PK136), Siglec-F (clone S17007L), FcγRIV (clone 9E9), Ly6C (clone HK1.4), CD3 (clone 17A2), CD19 (clone 6D5), CD11c (clone N418), CD45.2 (clone 104), Tim4 (clone F31-5G3), CD34 (clone HM34), CD16/32 (clone 93), CD135 (clone A2/F10), Sca-1 (clone D7), CD319 (clone 4G2), CD200R (clone OX-110), CD200 (clone OX-90), CD49e (clone 5H10-27), CD51 (clone RMV-7), CD22 (clone OX-97), Mac-2 (clone M3/38), CD14 (clone Sa14-2), and IL-6 (clone MP5-20F3) antibodies were purchased from BioLegend.
Monoclonal antibodies (mAbs) to mouse Ly6G (clone 1A8), B220 (clone RA3-6B2), CD115 (clone T38-320), CD117 (clone2B8), TER-119 (clone TER-119), and Grb2 (clone 81/GRB2) were purchased from BD biosciences (Franklin Lakes, NJ, USA). The mAbs anti-mouse MerTK (clone DS5MMER) and Tsc2 (clone SC05-59) were purchased from Invitrogen. The monoclonal anti-Tsc2 (clone D93F12) antibody was purchased from Cell Signaling Technology. The LEGENDScreen Mouse PE Kit was purchased from BioLegend.
Hiranuma R., Sato R., Yamaguchi K., Nakamizo S., Asano K., Shibata T., Fukui R., Furukawa Y., Kabashima K, & Miyake K. (2023). Aberrant monocytopoiesis drives granuloma development in sarcoidosis. International Immunology, 36(4), 183-196.
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4

Profiling Zbtb46+ Immune Cell Subsets

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ACK lysed splenocytes from three Zbtb46GFP/WT mice were isolated and enriched for CD19− CD3− cells. Cells were stained for 30 min at 4°C with the following 9 backbone antibodies and then washed: 7-AAD, SiglecH-AF647 (clone 551), CD11c-APC/Cy7 (clone N418), Bst2-PacBlue (clone 927), MHC-II-BV510 (clone M5/114.15.2), CD11b-BV785 (clone M1/70), XCR1-BV650 (clone ZET), CD19-Bio (clone 6D5), CD3-Bio (clone 17A2). The backbone labeled cells were then stained with 255 different PE-conjugated antibodies using the LegendScreen Mouse PE kit (BioLegend) following the manufacturer’s instructions. We used the Infinity Flow pipeline to combine overlapping flow cytometry panels and simultaneous analyze the co-expression patterns of hundreds of surface-expressed proteins across millions of individual cells as described in Becht et al. 54 (link). Clustering was performed using the Flowsom clustering method, with the parameters FlowSOM metacluster k = 7.
Rodrigues P.F., Kouklas A., Cvijetic G., Bouladoux N., Mitrovic M., Desai J.V., Lima-Junior D.S., Lionakis M.S., Belkaid Y., Ivanek R, & Tussiwand R. (2023). pDC-like cells are pre-DC2 and require KLF4 to control homeostatic CD4 T cells. Science immunology, 8(80), eadd4132.
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5

Spleen Leukocyte Screening for Cancer Markers

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A LEGENDScreen Mouse PE Kit (BioLegend) was used for spleen leukocyte screening for cancer-specific cell-surface markers. Plates from the kit were prepared according to the manufacturer’s instructions with lyophilized antibodies in each well of the assay plates being resuspended in 25 μL of deionized H2O (dH2O). The pooled barcoded and backbone antibody-labelled cells were added at 75 μL (i.e. 1x106 cell) to each well containing the reconstituted antibodies and incubated in the dark for 30 min at 4°C. Cells were then washed in Legend Screen Wash provided in the kit, pelleted and resuspended in 40 μL Labelling Buffer containing 1 μg/ml of the viability dye Hoechst 33285 (Invitrogen) and the equivalent of 500 Flow-Count Fluorospheres (7547053, Beckman Coulter) per 40 μL and stored at 4°C overnight before flow cytometry.
Simon Davis D.A., Ritchie M., Hammill D., Garrett J., Slater R.O., Otoo N., Orlov A., Gosling K., Price J., Yip D., Jung K., Syed F.M., Atmosukarto I.I, & Quah B.J. (2023). Identifying cancer-associated leukocyte profiles using high-resolution flow cytometry screening and machine learning. Frontiers in Immunology, 14, 1211064.
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