The following mAb were obtained from BD Biosciences (Heidelberg, Germany): anti-CD3-PE/APC/BV605 (clone SK7), anti-CD14-FITC/APC (clone MoP9), anti-PD-L1-BV421 (clone MIH1), and anti-IFN-γ-PE (clone 4S.B3). Anti-Vδ2-FITC (clone IMMU389) was obtained from Beckman Coulter (Krefeld, Germany), anti-CD277-PE (clone BT3.1) from BioLegend (San Diego, CA, USA). For cell surface staining, 3 × 105 cells were washed, stained for 20 min on ice with mAb, washed twice, and resuspended in 1% paraformaldehyde. For intracellular staining, cells were permeabilized in Cytofix/Cytoperm buffer (BD Biosciences) before staining with fluorochrome-conjugated mAb. All analyses were measured on a FACS Calibur or Fortessa cytometer (BD Biosciences), using the software Cell–Quest™ Pro and DIVA (Data-Interpolating Variational Analysis) for acquisition respectively, and FlowJo™ v10.6.1 software for data analysis.
Anti vδ2 fitc clone immu389
Manufactured by Beckman Coulter
Sourced in Germany
The Anti-Vδ2-FITC (clone IMMU389) is a fluorochrome-conjugated monoclonal antibody that binds to the Vδ2 chain of the T cell receptor. It is designed for use in flow cytometric analysis and can be used to identify and quantify Vδ2-positive T cells in biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Cytofix cytoperm buffer, by BD (2 mentions)
Lsr fortessa cytometer, by BD (2 mentions)
Anti ifn γ pe clone 4sb3, by BD (1 mentions)
Anti cd3 percp clone sk7, by BD (1 mentions)
Facscanto 2 flow cytometer, by BD (1 mentions)
Paraformaldehyde (pfa), by Merck Group (1 mentions)
Facscalibur, by BD (1 mentions)
Ptbk1 pe, by Cell Signaling Technology (1 mentions)
Cytofix cytoperm kit, by BD (1 mentions)
Facscanto, by BD (1 mentions)
4 protocols using anti vδ2 fitc clone immu389
1
Multicolor Flow Cytometry Protocol
2
Phenotypic Analysis of Vδ1 and Vδ2 T-cells
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Phenotype analysis of Vδ1 and Vδ2 T-cells was performed by using the following monoclonal antibodies: anti-Vδ2 FITC (clone IMMU389, Beckman Coulter Immunotech, Marseille, FR), anti-Vδ1 FITC (clone TS8.2; Thermo Scientific, USA), anti-CD3 PerCP (clone SK7, BD Pharmingen, San Jose, CA, USA), anti-CD27 APC (clone L128, BD Biosciences, San Jose, CA, USA), anti-CD45RA CY-Chrome (clone HI100, BD Biosciences San Jose, CA, USA), anti-CD3 AMCyan (clone SK7, BD Biosciences, Usa). The differentiation profile of Vδ1 and Vδ2 T-cells was analysed by monitoring the expression of CD45RA and CD27 markers. Specifically, Naïve was defined as CD45RA+CD27+, Central Memory as CD45RA-CD27+, Effector Memory as CD45RA-CD27-, and terminally differentiated as CD45RA+CD27-. Briefly, PBMC (1×106 cells/ml) were incubated with mAbs cocktail for 10 min a 4°C, washed once and fixed with 1% paraformaldehyde (1% PFA, Sigma, St. Louis, MS). Sample acquisition and data analysis were performed by a FACS Canto II Flow Cytometer (Becton Dickinson) by using Diva software.
3
Flow Cytometry Immunophenotyping Protocol
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The following mAb were obtained from BD Biosciences (Heidelberg, Germany): anti-CD3-PE/APC/BV605 (clone SK7), anti-CD14-FITC/APC (clone MoP9), anti-IFN-γ-PE or -PE/Cy7 (clone 4S.B3), anti-Granzyme B-PE-CF954 (clone GB11), anti-TNF-α-PE (clone MAb 11). Anti-Vδ2-FITC (clone IMMU389) was obtained from Beckman Coulter (Krefeld, Germany), anti-hTLR7-PE and anti-hTLR8-AF647 were purchased from R&D Systems. For cell surface staining, cells were washed, stained for 20 min on ice with mAb, washed twice, and resuspended in 1% paraformaldehyde. For intracellular staining, cells were permeabilized in Cytofix/Cytoperm buffer (BD Biosciences) before staining with fluorochrome-conjugated mAb. Final antibody concentrations were used according to respective data sheets. All analyses were measured on a FACS Calibur or LSR Fortessa cytometer (BD Biosciences), using the software CellQuest Pro and DIVA (Data-Interpolating Variational Analysis) for acquisition respectively, and FlowJo v10.6.1 software for data analysis.
4
Multiparametric Flow Cytometry Analysis
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The following mAb were obtained from BD Biosciences (Heidelberg, Germany): anti-CD3-PE/APC/BV605 (clone SK7), anti-CD14-FITC/APC (clone MoP9), anti-IFN-γ-PE or -PE/Cy7 (clone 4S.B3), anti-TNF-α-PE (clone MAb 11). Anti-STING-AF488 (clone 723505) was purchased from R&D Systems (Wiesbaden, Germany), anti-Vδ2-FITC (clone IMMU389) was obtained from Beckman Coulter (Krefeld, Germany), rabbit monoclonal pTBK1-PE (#13498S) from Cell Signaling Technology. For cell surface staining, 3 × 105 cells were washed, stained in V-bottom microtiter plates for 20 min on ice with mAb, washed twice, and resuspended in 1% paraformaldehyde. For intracellular staining, cells were washed with staining buffer and permeabilized using Cytofix/Cytoperm kit (BD Biosciences) before staining with fluorochrome-conjugated mAb. Staining for intracellular detection of pTBK1 in response to STING ligand stimulation was performed using the BD transcription factor buffer set. All analyses were measured on a FACS-Canto or LSR-Fortessa cytometer (BD Biosciences), using DIVA (Data-Interpolating Variational Analysis) for acquisition and FlowJo™ v10.6.1 for data analysis.
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