Truestain fcx plus

Manufactured by BioLegend

TrueStain FcX PLUS is a Fc receptor blocking reagent designed to reduce non-specific binding during flow cytometry experiments. It is formulated to effectively block Fc receptors on a variety of cell types, helping to improve the signal-to-noise ratio and accuracy of your data.

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Lab products found in correlation

Facsaria 3, by BD (1 mentions) Facslyric, by BD (1 mentions) Gentlemacs dissociator, by Miltenyi Biotec (1 mentions) Lung dissociation kit, by Miltenyi Biotec (1 mentions) Rbc lysis buffer, by BioLegend (1 mentions) Click it plus edu flow cytometry assay kit, by Thermo Fisher Scientific (1 mentions) Multi tissue dissociation kit 1, by Miltenyi Biotec (1 mentions) Percoll plus, by Cytiva (1 mentions) Live dead fixable near ir dead cell stain kit, by Thermo Fisher Scientific (1 mentions) Rneasy micro, by Qiagen (1 mentions) Smarter mouse tcr a b profiling kit, by Takara Bio (1 mentions) Miseq reagent kit v3, by Illumina (1 mentions) H 2kb mulv p15e tetramer, by MBL Life Science (1 mentions)

Spelling variants of this product

TrueStain fcX (21 citations) TrueStain FcX PLUS (anti-mouse CD16/32) antibody (3 citations)

2 protocols using truestain fcx plus

Single-cell isolation and flow cytometry

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Single cell suspensions were prepared from the bone marrow and spleen, followed by red blood cell (RBC) lysis by using RBC Lysis Buffer (BioLegend) for 5 min. For the flow cytometric analysis of peripheral blood, leukocytes from the heparinized blood sample were isolated from the interphase of 25% and 65% Percoll PLUS (cytiva) after gradient centrifugation. For the flow cytometric analysis of lung and skin samples, tissues were minced by razors and dissociated by using gentleMACS dissociator (Miltenyi Biotec) and Lung Dissociation Kit (Miltenyi Biotec, catalog #: 130-095-927) or Multi Tissue Dissociation Kit 1 (Miltenyi Biotec, catalog #: 130-110-201). Cells were stained with the indicated antibodies after treatment with normal rat serum (Merck Millipore) and TrueStain FcX PLUS (2.5 μg/mL; BioLegend; clone: S17011E, catalog#: 156604, dilution 1:200) to prevent non-specific binding. Stained cells were analyzed with FACSLyric (BD Biosciences) and FlowJo software ver 10.8.1 (BD Biosciences) or sorted with FACS AriaIII (BD Biosciences). For ex vivo EdU uptake assay, cells collected from the bone marrow or lung were cultured for 2 h with 10 μM EdU (5-ethynyl-2′-deoxyuridine). After culture, EdU staining was performed by using Click-iT Plus EdU flow cytometry assay kits (ThermoFisher Scientific, catalog #: C10633) according to the manufacturer’s protocol.

Miyake K., Ito J., Nakabayashi J., Shichino S., Ishiwata K, & Karasuyama H. (2023). Single cell transcriptomics clarifies the basophil differentiation trajectory and identifies pre-basophils upstream of mature basophils. Nature Communications, 14, 2694.

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Immune Profiling of Tumor-Infiltrating CD8+ T Cells

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Mice were inoculated with 2.0 × 10⁶ control LLC cells on the back (bilateral) and then treated with anti-PD1/CTLA4. On post challenge day 14–18, tumors were excised, weighed, and chopped before enzymatic and mechanical digestion, and immune cells were then isolated using density centrifugation and CD45⁺ magnetic bead enrichment, as described above. Cells were then stained in PBS using Live/Dead Fixable Near-IR Dead Cell Stain Kit (1:100, ThermoFischer), H-2Kb MuLV p15E Tetramer (1:10, MBL International), and TrueStain FcX Plus (anti-mouse CD16/32) Antibody (1:1000, BioLegend), followed by CD45 (1:100, Biolegend) and CD8 (1:10, MBL International). Samples were then pooled and analyzed using a MA900 FACS instrument (Sony), using single-colour compensation controls and fluorescence-minus-one controls for setting gates. Live/CD45⁺/CD8⁺/p15E⁺ cells were sorted into RMPI with 50% FBS. Live/CD45⁺/CD8⁺/p15E⁻ cells were also sorted. Cells were spun down and resuspended in RLT Buffer. RNA was subsequently isolated using RNeasy Micro (Qiagen). TCR’s were then isolated from the resulting RNA using the SMARTer Mouse TCR a/b Profiling Kit (Takara), per the manufacturer’s instructions. TCR’s were sequenced using a 600-cycle MiSeq Reagent Kit V3 for 2×300 base-pair reads (Illumina).

Griffin G.K., Wu J., Iracheta-Vellve A., Patti J.C., Hsu J., Davis T., Dele-Oni D., Du P.P., Halawi A., Ishizuka J.J., Kim S., Klaeger S., Knudsen N.H., Miller B.C., Nguyen T., Olander K., Papanastasiou M., Rachimi S., Robitschek E.J., Schneider E.M., Yeary M., Zimmer M.D., Jaffe J.D., Carr S.A., Doench J.G., Haining W.N., Yates K.B., Manguso R.T, & Bernstein B.E. (2021). Epigenetic Silencing by SETDB1 Suppresses Tumor Intrinsic Immunogenicity. Nature, 595(7866), 309-314.

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