Brilliant violet 421 anti

Manufactured by BioLegend

Sourced in United States

Brilliant Violet 421 anti-mouse CD45 is a fluorescently labeled antibody that binds to the CD45 surface antigen expressed on mouse leukocytes. It can be used for the identification and analysis of mouse immune cells by flow cytometry.

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2 protocols using brilliant violet 421 anti

Lung Cell Dissociation and Phenotyping

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Whole lungs were dissociated into single‐cell suspensions using the gentleMACS Dissociator. Red blood cell lysing buffer (Sigma‐Aldrich) was used for red cell lysis.
Lung cells were blocked with anti‐mouse CD16/32 (101319; Biolegend) and then stained with antibodies PerCP anti‐mouse/human CD11b (101229; Biolegend), Brilliant Violet 421 anti‐mouse F4/80 (123137; Biolegend), APC/Cy7 anti‐mouse CD45 (103116; Biolegend), PerCP/Cy5.5 anti‐mouse CD11c (117328; Biolegend), PE Siglec‐F (552126; BD Biosciences), PerCP/Cy5.5 anti‐mouse CD4 (100540; Biolegend), PE/Cy7 anti‐mouse CD3ε (100320; Biolegend), Brilliant Violet 421 anti‐mouse CD335 (NKp46) (137612; Biolegend), APC anti‐mouse CD8a (100712; Biolegend), Brilliant Violet 421 anti‐mouse Ly‐6G/Ly‐6C (Gr1) (108433; Biolegend) and Zombie Aqua Fixable Viability Kit (423102; Biolegend). Flow cytometric data acquisition was performed on BD FACS Canto II machine and data analysis was performed using FlowJo software.
Gating for CD45+AquaZombie−Siglec‐F+CD11c+Gr1− cells was used for AMs; CD45+AquaZombie−F4/80+CD11b+Gr1− for IMs CD45+AquaZombie−NKp46+ for natural killer (NK) cells, CD45+AquaZombie−CD3+CD8+ for CD8 T cells.

Alikhanyan K., Chen Y., Kraut S, & Sotillo R. (2020). Targeting alveolar macrophages shows better treatment response than deletion of interstitial macrophages in EGFR mutant lung adenocarcinoma. Immunity, Inflammation and Disease, 8(2), 181-187.

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Isolation and Characterization of Mammary Fibroblasts

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Single cell suspensions of normal mammary fat pads (n = 6) or 4T1 tumor-bearing BALB/c fat pads (n = 5) were generated as described above. Antibody staining was performed by incubating the cells with 1:100 dilution of APC/Cyanine7 anti-mouse I-A/I-E (MHC class II), Brilliant Violet 421™ anti-mouse CD90.2, PerCP/Cyanine5.5 anti-rat CD90/mouse CD90.1 (Thy-1.1), PerCP/Cyanine5.5 anti-mouse CD45, FITC anti-mouse Ly-6C (Biolegend, San Diego, CA, USA; Catalog no. 107627, 105341, 202515, 103131, 128005, respectively) for 30 min prior to resuspension in PBS with 1% FBS and 7-AAD Viability Staining Solution (BioLegend, San Diego, CA, USA; Catalog No. 420403). Cells were then analyzed on a BD (San Jose, CA, USA) FACSMelody instrument. Viable fibroblasts populations were identified as 7AAD−/CD45−/CD90.1−/CD90.2+ cells. Data analysis was performed using FlowJo software (v10.6.2).

Sebastian A., Hum N.R., Martin K.A., Gilmore S.F., Peran I., Byers S.W., Wheeler E.K., Coleman M.A, & Loots G.G. (2020). Single-Cell Transcriptomic Analysis of Tumor-Derived Fibroblasts and Normal Tissue-Resident Fibroblasts Reveals Fibroblast Heterogeneity in Breast Cancer. Cancers, 12(5), 1307.

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