Cell surface and intracellular staining was performed as previously described (15 (link), 47 (link)). Antibodies used: anti-CD3 Bv650 (OKT3, Biolegend), anti-CD3 FITC (SK7, BD Biosciences), anti-Vα7.2 PE (3C10, Biolegend), anti-CD161 Pe-Cy5 (DX12, BD Biosciences), anti-CD4 Bv711 (OKT4, Biolegend), anti-CD8 Bv570 (RPA-T8, Biolegend), anti-CD69 BUV737 (FN50, BD Biosciences), anti-CD107a BUV395 (H4A3, BD Biosciences), anti-GzB FITC (GB11, Biolegend), anti-IFNγ APC (25723.11, BD Biosciences), anti-TNF PE-Cy7 (Mab11, BD Biosciences), anti-IL17A Bv421 (BL168, Biolegend), anti-RORγt APC (R&D system), anti-RORγt Bv650 (Q21–559, BD Biosciences), anti-PLZF PECF594 (R17–809, BD Biosciences), anti-T-bet Bv711 or Bv605 (4B10, Biolegend), anti-MR1 (26.5, Biolegend), anti-HLA-DR (L243, Biolegend), anti-HLA-A2 (BB7.2, Biorad), LIVE/DEAD Fixable Aqua and Near-IR dye (Invitrogen). Flow cytometry data was acquired on BD LSRFortessa or BD Symphony A5 instruments (both BD Biosciences) and analyzed using FlowJo software v. 10.5.3 (TreeStar). Analyses of MAIT cell polyfunctionality were performed using the SPICE software v. 6.0 (48 (link)).
Anti rorγt bv650
Manufactured by BD
The Anti-RORγt Bv650 is a fluorescently-labeled antibody that binds to the RORγt (retinoic acid-related orphan receptor gamma t) transcription factor. RORγt is a key regulator of T helper 17 (Th17) cell differentiation and function. The Anti-RORγt Bv650 can be used in flow cytometry applications to detect and quantify RORγt-expressing cells.
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Lab products found in correlation
Anti ifn γ apc, by BD (1 mentions)
Anti cd3 fitc sk7, by BD (1 mentions)
Anti hla dr, by BioLegend (1 mentions)
Symphony a5, by BD (1 mentions)
Anti il 17a bv421, by BioLegend (1 mentions)
Anti mr1 26, by BioLegend (1 mentions)
Anti cd4 bv711 okt4, by BioLegend (1 mentions)
Anti cd107a buv395 h4a3, by BD (1 mentions)
Anti rorγt apc, by BD (1 mentions)
Anti plzf pe cf594, by BD (1 mentions)
Lsrfortessa, by BD (1 mentions)
Anti cd19 apc, by BD (1 mentions)
Anti cd3 apcr700, by BD (1 mentions)
Anti cd45ra bv 605, by BD (1 mentions)
Lsrfortessa x 20, by BD (1 mentions)
Anti foxp3 pe, by BD (1 mentions)
Anti cd45 buv395, by BD (1 mentions)
Anti cd16 pe cy7, by BD (1 mentions)
Anti cd25 bv421, by BD (1 mentions)
2 protocols using anti rorγt bv650
1
Multiparametric Flow Cytometry of MAIT Cells
2
Comprehensive Immune Cell Profiling
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To evaluate the relative proportions of lymphocyte populations in peripheral blood, the following panel of fluorochrome conjugated antibodies was used: anti CD45 BUV 395, anti CD3 APC R700, anti CD19 APC, anti CD16 PE-Cy7, anti CD56 BB515, anti CD4 BUV 7373, anti CD8 BV 785, anti CD25 BV 421, anti CD127 BB700, anti CD45RA BV605, anti CD197 BV711, anti FoxP3 PE, anti RORγT BV650 (all from BD Biosciences, Milan, Italy). Blood samples were stained with anti membrane associated molecules and, after incubation, erythrocytes were lysed using FACS lysing solution, according to manufacturer’s instruction. To analyze the expression of transcription factors (FoxP3 and RORγT), samples were fixed and permeabilized using Transcription Facytors Buffer Set (BD Bioscience), according to standard protocol. Samples were acquired and analyzed on a LSR Fortessa X-20 (Becton Dickinson), using FACS DIVA v8.0.2 software.
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