Fitc anti ly 6c clone al 21

Manufactured by BD

FITC-anti-Ly-6C (clone AL-21) is a fluorescently labeled antibody that binds to the Ly-6C antigen on the surface of cells. It is a tool for the identification and analysis of Ly-6C-expressing cells using flow cytometry or other applications.

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Lab products found in correlation

Phosflow fix buffer 1, by BD (2 mentions) Lsr 2, by FlowJo (2 mentions) Alexa fluor 594 anti vimentin clone epr3776, by Abcam (2 mentions) Pe cy5 labeled anti cd5 clone 53 7, by BioLegend (2 mentions) Pe cy7 anti gr 1 clone rb6 8c5, by Thermo Fisher Scientific (2 mentions) Pacific blue anti f4 80 clone bm8, by BioLegend (2 mentions) Perm buffer 3, by Thermo Fisher Scientific (2 mentions) Apc anti cd31 clone 390, by Thermo Fisher Scientific (2 mentions) Apc cy7 anti cd11b clone m1 70, by BioLegend (2 mentions) Anti cd16 32, by BD (2 mentions) Attune nxt cytometer, by Thermo Fisher Scientific (1 mentions) Percp cy5.5 anti cd11b clone m1 70, by BD (1 mentions) Anti gr1 pe clone rb6 8c5, by BD (1 mentions)

Spelling variants of this product

AL-21 (6 citations) Clone AL-21 (4 citations)

3 protocols using fitc anti ly 6c clone al 21

Multiparametric Flow Cytometry of Aortic and Splenic Immune Cells

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Single cell suspensions from the aortae and spleen were obtained as described above and incubated with anti-CD16/32 (BD Biosciences, 553142) and stained on ice for 30 minutes with the following antibodies: Alexa Fluor 594-anti-Vimentin (clone EPR3776, Abcam, ab154207), APC-anti-CD31 (clone 390, Invitrogen, 17–0311-80), FITC-anti-Ly-6C (clone AL-21, BD Biosciences, 553104), PE-Cy5-labeled anti-CD5 (clone 53–7.3, BioLegend, 100609), PE-Cy7-anti-Gr-1 (clone RB6–8C5, Invitrogen, 25–5931-81), APC-Cy7-anti-CD11b (clone M1/70, BioLegend, 101225), and Pacific Blue-anti-F4/80 (clone BM8, BioLegend, 123123). For intracellular staining, cells were fixed and permeabilized with buffers (BD Phosflow Fix Buffer I and Perm Buffer III) according to the manufacturer’s instructions, then stained with Alexa Fluor 488-anti-alpha-smooth muscle actin (α-SMA, clone 1A4, eBioscience, 50–112-4644). Cell suspensions were subjected to flow cytometry (Becton Dickinson LSR II) and analyzed using FlowJo10.1.r5. Macrophages were identified as CD11b⁺/Ly-6C^low/F4/80⁺ cells. Ly-6C^hi monocytes were identified as CD11b⁺/Ly-6C^hi/F4/80^low cells. Neutrophils were identified as CD11b⁺/Gr-^hi cells.

Flores A.M., Hosseini-Nassab N., Jarr K.U., Ye J., Zhu X., Wirka R., Koh A.L., Tsantilas P., Wang Y., Nanda V., Kojima Y., Zeng Y., Lotfi M., Sinclair R., Weissman I.L., Ingelsson E., Smith B.R, & Leeper N.J. (2020). Pro-efferocytic nanoparticles are specifically taken up by lesional macrophages and prevent atherosclerosis. Nature nanotechnology, 15(2), 154-161.

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Multiparametric Flow Cytometry of Aortic and Splenic Immune Cells

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Treg Cell and MDSC Quantification

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The CD4+Foxp3+ (Treg cells) and MDSC were measured by flow cytometry at day 7 post immunization protocol and at day 21 post infection. Briefly, mice were sacrificed and spleen single cell suspensions were prepared by homogenization through a stainless steel mesh using PBS 3% fetal bovine serum (FBS). Spleen absolute numbers were determined by counting cells in Neubauer chamber using Turk solution and then red blood cells were eliminated by lysis with H₂0 during 30 seconds. Finally, spleen cells were homogenized in PBS 3% FBS and 1.10⁶ cells were stained with the following antibodies from BD-Pharmingen: fluorescein isothiocyanate (FITC) anti-CD4 (clone RM4-5), FITC anti-Ly6C (clone AL-21), phycoeritrin (PE) anti-Ly6G (clone 1A8), PE anti-GR1 (clone RB6-8C5) and PerCP-Cy 5.5 anti-CD11b (clone M1/70). Cells marked with CD4 were also intracellular stained for PE anti-Foxp3, using mAb from Miltenyi Biotec, according to manufacturer´s instructions.
Samples were acquired on an Attune NxT cytometer (Invitrogen) and analyses were performed using FlowJo software.

Prochetto E., Roldán C., Bontempi I.A., Bertona D., Peverengo L., Vicco M.H., Rodeles L.M., Pérez A.R., Marcipar I.S, & Cabrera G. (2017). Trans-sialidase-based vaccine candidate protects against Trypanosoma cruzi infection, not only inducing an effector immune response but also affecting cells with regulatory/suppressor phenotype. Oncotarget, 8(35), 58003-58020.

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