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Cd45 apc cy7 30 f11

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CD45-APC/Cy7 (30-F11) is a fluorescent-conjugated antibody that binds to the CD45 antigen, a common leukocyte antigen expressed on the surface of various immune cells. This product is designed for use in flow cytometry applications to identify and analyze different populations of leukocytes.

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Lab products found in correlation

Live dead fixable aqua dead cell stain kit, by Thermo Fisher Scientific (1 mentions) Cd16 cd32 fc block, by BioLegend (1 mentions) Ly6c apc hk1, by BioLegend (1 mentions) Cd11c pe cy7, by BioLegend (1 mentions) Cytoflex lx, by Beckman Coulter (1 mentions) Dnase 1, by Merck Group (1 mentions) Liberase tl, by Roche (1 mentions) Efluor 506, by Thermo Fisher Scientific (1 mentions) Cd3 percp cy5.5 145 2c11, by BioLegend (1 mentions) Ly6 c pe cy7 hk1.4, by BioLegend (1 mentions) Cd11b percp cy5.5 m1 70, by BioLegend (1 mentions) Gentlemacs dissociator, by Miltenyi Biotec (1 mentions) Ghost violet 510 viability dye, by Cytek Biosciences (1 mentions) Fc block 2.4g2, by BD (1 mentions) Cd8 percpcy5.5 53 6 7, by BioLegend (1 mentions) Cd4 bv785 gk1.5, by BioLegend (1 mentions) Fixation and intracellular staining permeabilization wash buffer, by BioLegend (1 mentions) Facscanto 2, by Tree Star (1 mentions) Ack lysing buffer, by Thermo Fisher Scientific (1 mentions) Live dead aqua, by Thermo Fisher Scientific (1 mentions)

4 protocols using cd45 apc cy7 30 f11

1

Neutrophil Isolation from Murine Skin

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10-mm skin punch biopsies were minced and digested in 3 mL RPMI containing 100 μg/mL DNase I (Sigma-Aldrich) and 1.67 Wunsch units/mL Liberase TL (Roche) for one hour at 37 °C on a rotor wheel. Single cell suspensions were generated by passing digested skin samples through 100-μm cell strainers and washed in RPMI and PBS. Live versus Dead cells were stained using Live/Dead Fixable Aqua Dead Cell Stain Kit (Thermo Fisher). Cells were treated with CD16/CD32 Fc Block (S17011E, Biolegend) for 10 min before incubation with the antibody cocktail: CD45-APC/Cy7 (30-F11, Biolegend), CD11b-PE/Dazzle594 (M1/70, Biolegend), CD11c-PE/Cy7 (N418, Biolegend), Ly6C-APC (HK1.4, Biolegend), Ly6G-BV421 (1A8, Biolegend), F4/80-BV605 (BM8, Biolegend) and SiglecF-BB515 (E50-2440, BD Bioscience). Data were collected using a CytoFLEX LX (Beckman Coulter) and analyzed using FlowJo (BD). Neutrophils were gated as live CD45+CD11b+Ly6G+.
Dong X., Limjunyawong N., Sypek E.I., Wang G., Ortines R.V., Youn C., Alphonse M.P., Dikeman D., Wang Y., Lay M., Kothari R., Vasavda C., Pundir P., Goff L., Miller L.S., Lu W., Garza L.A., Kim B.S., Archer N.K, & Dong X. (2022). Keratinocyte-derived defensins activate neutrophil-specific receptors Mrgpra2a/b to prevent skin dysbiosis and bacterial infection. Immunity.
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2

Isolation and Characterization of Nonparenchymal Cells from Mouse Liver

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Nonparenchymal cells were isolated from mouse livers using a gentleMACS dissociator (Miltenyi Biotec, Bergisch Gladbach, Germany) and following the protocol for the preparation of single-cell suspensions from mouse liver provided by the manufacturer. Nonparenchymal cells were stained for different cell surface markers based on a previously described protocol.58 (link) The following antibodies (Biolegend, San Diego, CA, USA) were used: CD45-APC-Cy7 (30-F11), CD3-PerCP-Cy5.5 (145-2C11), CD4-Brilliant Violet 421 (GK1.5), CD8-PE-Cy7 (53-6.7), CD19-APC (6D5), CD11b-PerCP-Cy5.5 (M1/70), CD11 c-APC (N418), F4/80-PE (BM8) and Ly6 c-PE-Cy7 (HK1.4). For dead/live discrimination eFluor® 506 (eBioscience, San Diego, CA, USA) was used and cell suspensions were analyzed by flow cytometry. The gating strategy is illustrated in Supplementary Fig. 11.
Foerster F., Boegel S., Heck R., Pickert G., Rüssel N., Rosigkeit S., Bros M., Strobl S., Kaps L., Aslam M., Diken M., Castle J., Sahin U., Tuettenberg A., Bockamp E, & Schuppan D. (2017). Enhanced protection of C57 BL/6 vs Balb/c mice to melanoma liver metastasis is mediated by NK cells. Oncoimmunology, 7(4), e1409929.
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3

Comprehensive Immune Cell Phenotyping

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Cells isolated from mouse spleen and thymus were stained with Ghost Violet™ 510 viability dye (TONBO Biosciences) for dead cell exclusion, blocked with Fc block (2.4G2, BD Biosciences) and stained with the following anti-mouse antibodies from Biolegend: CD4-BV785 (GK1.5), CD44-PE-Cy7 (IM7), TCRb-APC (H57-597), CD8-PerCP-Cy5.5 (53-6.7) and CD45-APC-Cy7 (30-F11). Cells were then fixed and permeabilized using fixation and intracellular staining permeabilization wash buffer from Biolegend following the manufacturer’s instructions. Cells were stained with mouse anti-IRAP-AF594 (F5, Santa Cruz Biotechnology, dilution 1/20) or with rabbit a-Stx6 (ProteinTech, dilution 1/100) or monoclonal rabbit IgG (Cell Signaling Technology, dilution 1/500) followed by staining with a-rabbit AF594 (Thermo Fisher Scientific, dilution 1/200).
Evnouchidou I., Chappert P., Benadda S., Zucchetti A., Weimershaus M., Bens M., Caillens V., Koumantou D., Lotersztajn S., van Endert P., Davoust J., Guermonprez P., Hivroz C., Gross D.A, & Saveanu L. (2020). IRAP-dependent endosomal T cell receptor signalling is essential for T cell responses. Nature Communications, 11, 2779.
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4

Lung Leukocyte Isolation and Macrophage Phenotyping

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Lung leukocytes were isolated by digesting matched lung lobes in DNase (MilliporeSigma) (25 μg/mL) and Liberase (MilliporeSigma) (18 μg/mL) for 60 minutes with rocking at 37°C, crushing through 70-μm cell strainers, and centrifuging at 350 g for 10 minutes. Erythrocytes were eliminated using ACK Lysing Buffer (Gibco). Lung leukocyte numbers were determined by flow rate. To assess cell populations and macrophage phenotype, lung leukocytes (4 × 105) were labeled with a viability dye (Invitrogen Live/Dead Aqua), preincubated with unlabeled anti-CD16/32 antibody, and stained with: CD45-APC-Cy7 (30-F11, BioLegend), CD11b-Percp (M1/70, BioLegend), F4/80-BV421 (BM8, BioLegend), CD206-APC (C068C2, BioLegend), and iNOS (K13-A, Biorbyt). Macrophages were gated as CD45+F4/80+/CD11b+ and then separated into distinct M1 (CD45+F4/80+/CD11b+/iNOS+) and M2 (CD45+F4/80+/CD11b+/CD206+) subsets. Flow cytometry was performed using a FACSCanto II and data were analyzed using FlowJo (Tree Star).
Leslie J., Millar B.J., del Carpio Pons A., Burgoyne R.A., Frost J.D., Barksby B.S., Luli S., Scott J., Simpson A.J., Gauldie J., Murray L.A., Finch D.K., Carruthers A.M., Ferguson J., Sleeman M.A., Rider D., Howarth R., Fox C., Oakley F., Fisher A.J., Mann D.A, & Borthwick L.A. (2020). FPR-1 is an important regulator of neutrophil recruitment and a tissue-specific driver of pulmonary fibrosis. JCI Insight, 5(4), e125937.
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