Brilliant Violet 785™ anti-mouse CD117 (c-kit) clone 2B, Brilliant Violet 785™ anti-mouse CD117 (c-kit) clone 2B8, Brilliant Violet 421™ anti-mouse Ly-6A/E (Sca-1) Antibody Clone D7 BV421 (BioLegend); CD135 (Flt3) Monoclonal Antibody (A2F10), PerCP-eFluor 710, PerCP-eFluor 710 (eBioscience); PE Rat anti-Mouse CD34 Clone RAM34 (RUO), Ms. CD16/CD32 BV605 2.4G2 and Ms. CD127 BUV737 SB/199 (BD Biosciences).
Percp efluor 710
Manufactured by Thermo Fisher Scientific
Sourced in United States
PerCP-eFluor 710 is a fluorescent dye used in flow cytometry applications. It is a tandem dye, composed of the PerCP (Peridinin-Chlorophyll Protein Complex) and eFluor 710 fluorophores. The PerCP component absorbs light and transfers energy to the eFluor 710 fluorophore, which then emits a detectable signal. This dye is often used to label and detect cell surface markers in multi-color flow cytometry experiments.
Automatically generated - may contain errors
Lab products found in correlation
Attune nxt, by Thermo Fisher Scientific (6 mentions)
Apc efluor 780, by Thermo Fisher Scientific (5 mentions)
Fixable viability dye efluor 780, by Thermo Fisher Scientific (2 mentions)
Apc fire 750, by BioLegend (2 mentions)
Fluorescein isothiocyanate (fitc), by Thermo Fisher Scientific (2 mentions)
Efluor 450, by Thermo Fisher Scientific (2 mentions)
Triton x 100, by Merck Group (2 mentions)
Propidium iodide, by Merck Group (2 mentions)
Hit3a, by BioLegend (1 mentions)
Mca1774f, by Bio-Rad (1 mentions)
Fixable viability dye 780, by Thermo Fisher Scientific (1 mentions)
Cytexpert software, by Beckman Coulter (1 mentions)
Cytoflex s flow cytometer, by Beckman Coulter (1 mentions)
Stereomicroscope, by Zeiss (1 mentions)
Alexa fluor 647, by Thermo Fisher Scientific (1 mentions)
Ms 295 p1, by Thermo Fisher Scientific (1 mentions)
Saponin, by Merck Group (1 mentions)
Sh800 cell sorter, by Sony (1 mentions)
Mbsa43, by Thermo Fisher Scientific (1 mentions)
Live dead cell viability assay, by Thermo Fisher Scientific (1 mentions)
33 protocols using percp efluor 710
1
Multiparametric Analysis of Mouse Hematopoietic Stem Cells
2
Comprehensive Immunophenotyping of CD8+ T Cells
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Flow cytometry of CD8+ T cells was performed using a CytoFLEXS FACS (Beckman Coulter Life Sciences, Mississauga, Ontario, Canada) to detect IRs, cytokine levels and proliferation. Lymphocytes were stained with 0.2 µg of each of the following MAbs: FITC anti-mouse CD8, PerCP-eFluor 710 anti-mouse PD-1, PE-CY7 anti-mouse TIM-3, APC anti-mouse BTLA, PerCP-eFluor 710 Rat IgG2b Isotype Control, PE Mouse IgG2a κ Isotype Control, FITC Mouse IgG2a κ Isotype Control, PE-CY7 Mouse IgG2a κ Isotype Control, APC Mouse IgG1 κ Isotype Control, PE anti-mouse IL-2 PE anti-mouse TNF-α and PE anti-mouse IFN-γ- (eBioscience, San Diego, CA, USA). A Fixation/Permeabilization Solution Kit (BD Biosciences, USA) was used for intracellular staining where required for different experiments.
3
Flow Cytometric Analysis of Monocyte Markers
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The expression of specific markers was investigated in monocytes from synovial fluid by fluorescence-activated cell sorting analysis after surface or intracellular staining with specific monobodies that were conjugated to different fluorescent dyes. For the extracellular staining, cells were washed in phosphate-buffered saline (PBS) containing 1% bovine serum albumin and 0.02% sodium azide and were incubated with specific antibodies for 30 min at 4°C. Flow cytometry was performed using a FACS Canto (CytoFLEX; Beckman Coulter, CA, USA) and analyzed with Flowjo analysis software (Tree Star, Inc., Ashland, OR, USA). Specific monoclonal antibodies to human CD11c (Alexa Fluor® 700; cat. no. 56-0116-42), CD206 (Alexa Fluor® 488; cat. no. 53-2069-42), CD86 (Alexa Fluor® 488; cat. no. A51007) and CD163 (PerCP-eFluor® 710; cat. no. 46-1639-42) were all from Thermo Fisher Scientific, Inc. Staining with mouse IgG isotype control (PerCP-eFluor® 710 labeled, cat. no. 46-4714; Thermo Fisher Scientific, Inc.) was used as control for analysis of monocytes in the monocyte gate.
4
Phenotypic Characterization of EMD Cells
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EMD cells were thawed in RPMI supplemented with 10% FCS. Staining was performed in PBS at 4 °C with the following antibodies: CD184/CXCR4 – PerCP/eFluor710 (1:100, 12G5, Thermofisher), CD326/EpCAM – APC (1:100, 9C4, Biolegend), CD269/BCMA – PE (1:100, REA361, Miltenyi), CD38- FITC (1:100, HIT2, Thermofisher), CD138 – BV510 (1:20, MI15, Biolegend), CD45 – BV605 (1:20, 2D1, Biolegend), CD3 – AF700 (1:100, HIT3a, Biolegend). For dead cell exclusion, Fixable Viability Dye eFluor ™ 780 (1:1000, Thermofisher) was used. A FcGr blocking step was not performed. Events were acquired on the Attune NxT (Thermofisher). Data was analyzed with FlowJo version 10.8.0.
5
Multiparametric Immune Cell Analysis
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CD3-FITC (clone CA17.2A12, BioRad #MCA1774F), NKp46-PE (clone 48A, kind gift of Dr. Dean Lee), CD5 on PerCP-eFluor 710 (clone YKIX322.3) Thermo Fisher #46-5050-42, Live/dead staining performed using Fixable Viability Dye 780 (eBioscience #65-0865-14).
6
Multiparameter Flow Cytometry Immunophenotyping
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In brief, 50 μL of EDTA‐anticoagulated whole blood was stained with saturating concentrations of the following antibodies: CD11b/c (PerCP‐eFluor 710, Thermo Fisher Scientific, Massachusetts, USA), anti‐granulocyte marker (FITC, eBioscience, California, USA), and CD43 (Alexa Fluor 647, BioLegend, California, USA) following standard protocols on an Attune NXT (Thermo Fisher). The entire gating strategy is shown in FigureS2 .
7
Apoptosis and Cell Cycle Analysis
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For assessment of apoptosis, cells were first seeded into 6-well tissue culture plates to give approximately 50% confluence and allowed to attach overnight. The cells were then treated for 24 h with the indicated compound. Apoptosis was evaluated using an Annexin V apoptosis detection kit PerCP-eFluor™ 710 (cat#88-8008, Thermo Fisher Scientific, Waltham, MA, USA). Data was acquired using a CytoFLEX S flow cytometer (Beckman Coulter, Brea, CA, USA) comprising 10,000 events on the PC5.5 channel and analyzed with CytExpert software (Beckman Coulter). Cell cycle analysis was performed by ethanol fixation of cells, followed by incubation with 1 µg/mL Hoechst 33258.
8
Embryonic Stem Cell Characterization
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Mouse embryos (E7.75) were dissected using forceps under a stereomicroscope (Zeiss) and regions of interest were dissociated and harvested using TrypLE. Embryoid bodies (EBs) and cells were dissociated and harvested using TrypLE. Single-cells were analyzed for RFP/GFP expression or sorted using a SH800 Cell sorter (Sony Biotechnologies). Live cells were analyzed for RFP and GFP expression and stained with antibodies targeting for the presence of appropriate markers. Cells were stained with the following antibodies: anti-mouse Cxcr4 conjugated with PerCP-eFluor 710 (1:200; 46-9991-80 eBiosciences) anti-mouse EphA2 conjugated with APC (1:100; Cat. FAB639A R&D systems), anti-human Cxcr4 conjugated with PE or APC (1:25; Cat. FAB170P R&D systems). For cTNT and Isl1 expression, cells were fixed with 4% paraformaldehyde (PFA) for 10 min, permeabilized with saponin (Sigma), stained with either mouse cTNT (1:500, Cat. MS-295-P1 Thermo Scientific) or mouse Islet1 antibody (1:200, Cat. 39.3F7 Developmental Studies Hybridoma Bank, Iowa City, IA), followed by incubation with secondary antibody conjugated with Alexa Fluor 647 (1:500, Invitrogen). Data were analyzed using FlowJo software.
9
Multiparametric Analysis of Tumor-Infiltrating T Cells
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Peripheral blood mononuclear cells were isolated from whole blood by Ficoll-Paque gradient centrifugation. Freshly resected tumor specimens were manually disrupted followed by digestion with collagenase IV (2.5 mg/mL) and DNase I (0.2 mg/mL) (Worthington Biochemical Corporation) for 1 hour. Tumor homogenates were separated on discontinuous 70%–30% Percoll (Sigma-Aldrich) gradients. Flow cytometric analysis was performed with antibodies targeting CD4 (BD Biosciences clone RPA-T4, V450 conjugated), CD8 (BD Biosciences clone RPA-T8, V500 conjugated), TIGIT (eBioscience clone MBSA43, PerCP-eFluor® 710 conjugated), CD226 (eBioscience clone TX25, FITC conjugated), and PD-1 (BD Biosciences clone EH12.1, Alexa Fluor® 647 conjugated). Cell viability was assessed using Live/Dead Cell Viability Assays (Life Technologies). Samples were run on a BD LSRFortessa or BD FACSAria II, as previously described. FlowJo software (Tree Star Inc.) was used for analysis after gating on live cells, with doublet exclusion followed by gating on CD4 and CD8 T cells.
10
Multiparameter Flow Cytometry Assay
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Antibodies against murine CD4 (RM4-5, AF700, BV650), CD8 (53-6.7, APC/Fire 750, BV785), CCR2 (SA203G11, BV421), CD11c (N418, APCCy7, Pe/Cy5), CD206 (C068C2, APC), CD11b (M1/70, BV421), F4/80 (BM8, BV510, BV711, FITC), TCRβ (H57-597, BV605), and NK1.1 (PK136, BV650) were purchased from BioLegend. Antibodies against CD4 (GK1.5, BUV395) and ST2 (U29-93, BV480) were purchased from BD Biosciences. Antibodies against B220 (RA3-6B2, AF488), CD11c (N418, e450, PE-Cy5.5), CD25 (PC61.5, PE-Cy7), CD4 (GK1.5, APC-eFluor 780, Super Bright 645), CD45 (30-F11, Pacific Orange), Foxp3 (FJK-16s, APC, e450, FITC), GITR (DTA-1, Super Bright 600), IL-10 (JES5-16E3, PE), KLRG1 (2F1, APC, APC-eFluor 780, PE-eFluor 610), NK1.1 (PK136, PE), and ST2 (RMST2-2, PE, PerCP-eFluor710) were purchased from eBioscience, Thermo Fisher Scientific. Viability dyes (e506, near IR, UV) were purchased from Invitrogen, Thermo Fisher Scientific.
Extracellular flow staining was performed in 2% FBS-PBS in the presence of BioLegend’s TruStain FcX for 30 minutes at 4°C. Fixation, permeabilization, and intracellular staining were performed using eBioscience’s (Thermo Fisher Scientific) Foxp3/transcription factor staining buffer set per manufacturer’s directions. Flow data were collected using a Cytek Aurora or BD Biosciences LSR and analyzed using FlowJo.
Extracellular flow staining was performed in 2% FBS-PBS in the presence of BioLegend’s TruStain FcX for 30 minutes at 4°C. Fixation, permeabilization, and intracellular staining were performed using eBioscience’s (Thermo Fisher Scientific) Foxp3/transcription factor staining buffer set per manufacturer’s directions. Flow data were collected using a Cytek Aurora or BD Biosciences LSR and analyzed using FlowJo.
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