Fluorescein isothiocyanate (FITC) anti-CD3 (SK7); FITC anti-CD19 (HIB19); FITC anti-CD28 (CD28.2); FITC anti-CD45RO (UCHL1); FITC anti-CD56 (NCAM16.2); FITC anti-CD69 (FN50); FITC anti-CD94 (HP-3D9); FITC anti-CD95 (DX2); FITC anti-TCR αβ (WT31); phycoerythrin (PE) anti-CD3 (SK7); PE anti-CD25 (M-A251); PE anti-CD45RA (HI100); PE anti-CD56 (NCAM6,2), PE anti-IL-2 (MQ1-17H12); 7-Amino-Actinomycin D (7-AAD); PE-Cy5 anti-CD3 (UCHT1); PE-Cy7 anti-CD3 (SK7); PE-Cy7 anti-CCR7 (3D12); PE-Cy7 anti-IFNγ (B27); allophycocyanin (APC) anti-CD3 (SK7); APC anti-CD4 (RPA-T4); APC anti-CD8 (RPA-T8); APC anti-CD27 (L128); APC anti-CD45RO (UCHL1); APC-Cy7 anti-CD8 (SK1); Alexa Fluor 700 anti-CD4 (RPA-T4); V450 anti-CD3 (UCHT1) were purchased from BD Biosciences, San Jose, CA, USA. FITC anti-FOXP3 (236A/E7); APC anti-FOXP3 (236A/E7) and Alexa Fluor 700 anti-CD4 (OKT-4) came from eBioscience, Inc., San Diego, CA, USA. Qdot605 anti-CD3 (UCHT1) and Pacific Orange anti-CD8 (3B5) came from Invitrogen, Eugene, OR, USA. FITC anti-TCR PANγδ (IMMU510) and Krome Orange anti-CD4 (13B8.2) was purchased from Beckman Coulter, Immunotech, Marseille, France.
Anti cd3 pe cy5
Manufactured by BD
Sourced in United States, United Kingdom
Anti-CD3-PE-Cy5 is a fluorochrome-conjugated monoclonal antibody that binds to the CD3 receptor complex on the surface of T cells. It can be used for the identification and enumeration of T cells in flow cytometry applications.
Automatically generated - may contain errors
Lab products found in correlation
Anti cd4 fitc, by BD (8 mentions)
Anti cd19 fitc, by BD (3 mentions)
Anti cd8 pe, by BD (3 mentions)
Anti cd19 pe, by BD (3 mentions)
Anti cd56 pe cy7, by BD (3 mentions)
Anti cd8 pe cf594, by BD (3 mentions)
Facscalibur flow cytometer, by BD (3 mentions)
Anti cd56 pe, by BD (2 mentions)
Facs lysing solution, by BD (2 mentions)
Anti cd25 pe cy5, by BD (2 mentions)
Anti cd3 fitc anti cd16 pe anti cd56 pe, by BD (2 mentions)
Cellquest pro software, by BD (2 mentions)
Anti cd3 fitc, by BD (2 mentions)
Cxp analysis software, by Beckman Coulter (2 mentions)
Cytomics fc500 flow cytometer, by Beckman Coulter (2 mentions)
Cyan adp 9 colour flow cytometer, by Beckman Coulter (2 mentions)
Anti cd56 fitc, by BD (1 mentions)
Anti cd69 fitc, by BD (1 mentions)
Anti cd28 fitc, by BD (1 mentions)
7 aminoactinomycin d, by BD (1 mentions)
16 protocols using anti cd3 pe cy5
1
Multiparametric Flow Cytometry Panel
2
Molecular Mechanisms of Inflammation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The oxazolone and sulfasalazine were purchased Sigma-Aldrich (St. Louis, MA, USA). Primary antibodies for western blot including phosphorylated (p)–NF–κB (Catalog No. 3033), and NLRP3 (Catalog No. 15101), and horseradish peroxidase-linked secondary antibody (Catalog No. 7074) were purchased Cell Signaling Technology (Danvers, MA, USA). The conjugated fluorescence antibodies for flow cytometry analysis (FACS) including PE-Cy5 anti-CD3, PE anti-CD4, fluorescein isothiocyanate (FITC) anti-CD8, FITC anti-CD19, and FITC anti-CD25, were obtained BD Biosciences (San Diego, CA, USA). All other chemicals and reagents were obtained from Sigma Aldrich.
3
Immunophenotyping Peripheral Blood Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Samples for cytometric analyses were prepared from freshly obtained peripheral blood incubated with a set of monoclonal antibodies: anti-CD3 FITC, anti-CD3 PECy5, anti-CD4 FITC, anti-CD8 PE, anti-CD19 PE, anti-iNKT FITC, anti-CD25 PECy5, and anti-CD3 FITC/anti-CD16 PE/anti-CD56PE (BD Pharmingen, USA). The samples were deprived of erythrocytes by adding lysing solution (FACS Lysing Solution, Becton Dickinson, USA). The immunophenotype of peripheral blood cells was determined with a FACSCalibur flow cytometer (Becton Dickinson, USA) equipped with an argon laser emitting at 488 nm. The results were analyzed with CellQuest Pro software (Becton Dickinson, USA).
Statistical analysis was carried out with the R Project for Statistical Computing v. 3.4. Descriptive statistics were calculated. The Student’s t test, F test, and Brown–Forsythe test were used for comparison between groups of variables. Associations between percentages and numbers of NK, NKT, CD3, and CD3CD8 cells in patients with recurrent furunculosis were assessed with Pearson coefficient for dependent variables. The criteria for statistical significance were set at p < 0.05.
Statistical analysis was carried out with the R Project for Statistical Computing v. 3.4. Descriptive statistics were calculated. The Student’s t test, F test, and Brown–Forsythe test were used for comparison between groups of variables. Associations between percentages and numbers of NK, NKT, CD3, and CD3CD8 cells in patients with recurrent furunculosis were assessed with Pearson coefficient for dependent variables. The criteria for statistical significance were set at p < 0.05.
4
Flow Cytometric Analysis of Immune Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
This was undertaken according to our previously published methods (33 (link)). The method utilized whole blood analysis with red cell lysis using Easylyse (BD Biosciences) and with anti-CD56 PE, anti-CD16 FITC, anti-CD3 PE Cy5 and anti-CD69 APC (all supplied by BD Pharmingen). Flow cytometric analysis was undertaken on a Facscalibur (BD Biosciences), and using Cellquest Pro software. A minimum number of 10,000 cells were counted.
5
Differentiation of Th Subsets from CD4+ T Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
CD4+ T cells isolation from PBMCs lymphocytes were performed using human CD4 T Cell isolation kit (Miltenyi Biotec, Marburg, Germany) according to the manufacturer’s protocol. CD4+ T cells were stimulated with of anti-CD3/CD28 microbeads and IL-2 (20 ng/mL) and added with specific cytokine for the differentiation of specific Th cells; IFNγ (25 ng/mL) and IL-12 (25 ng/mL) for Th1; IL-6 (50 ng/mL) and TGFβ (25 ng/mL) for Th17; TGFβ (25 ng/mL) and Retinoic acid (10 nM) for Treg. Cytokine-treated CD4+ T cells (5 × 105/well) were cultured for 5 days by adding EVs derived from naive or TSG primed WJ-MSCs in a 24 well plate. For intracellular staining, cells were stimulated with a cell stimulation cocktail (Invitrogen) for 5 h before staining. The cells were stained with cell surface marker anti-CD3-PE-Cy5, anti-CD4-FITC and anti-CD25-APC (clone M-A251, BD Biosciences) antibodies and then fixed/permeabilized and stained with fluorescence-labeled anti-IFNγ-APC (clone 4S.B3), anti-IL-17A-PE (clone SCPL1362), and anti-Foxp3-PE (clone 259D/C7, BD Biosciences) antibodies. Staining cells were analyzed by flow cytometry.
6
Immune Response Analysis of RVFV Gn Protein
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
T cell subset populations and memory T lymphocytes in splenocytes were analyzed by flow cytometry. Splenocytes were seeded into six-well plates at a density of 5 × 106 cells/mL and stimulated with or without purified RVFV Gn head protein (10 μg/mL) for 36 h at 37°C and 5% CO2. Cells were then labeled with equal volumes of 1:250 dilutions of anti-CD3-PE-Cy5, anti-CD4-FITC, anti-CD8-PE, anti-CD44-APC, and anti-CD62-PerCP-Cy5 (BD Biosciences, United States) for 30 min at 4°C. After washing, labeled cells were analyzed in a FACSAria™ Cell Sorter (BD Biosciences, United States).
7
Phenotypic Characterization of Activated T Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
PBMC were purified as indicated above and activated with TransAct™ (1:100, Miltenyi-Biotec 130-111-160) and recombinant human IL2 (100 IU/ml; PeproTech). Then at days 0 and days 2-4-6 post-stimulation, the cells were stained with Fixable Viability Stain 450 (250 ng/mL; BD Biosciences), next with anti-CD3-PE-Cy5 (1/30; clone HIT3a; BD Biosciences), anti-CD4-PE-Vio770 (1/100; clone rea623; Miltenyi Biotec), anti-CD8-APC-Cy7 (1/30; clone RPA-T8; Biolegend), anti-CD25-Alexa fluor 488 (1/30; clone M-A251; Biolegend), anti-CD45RA-APC (1/30; clone HI100; Biolegend), anti-CCR7-PE (1/50; clone G043H7; Biolegend), anti-Granzyme-B-PE (1/50; clone GB12; Invitrogen), anti-PD1-Alexa 488 (1/50; clone EH12-2H7; Biolegend) and anti-KLRG1-APC (1/100; clone Rea261; Miltenyi Biotec). FACS analyses were performed on a MACSQuant Analyzer (Miltenyi Biotec) and data analyzed using FlowJo 10 software (FlowJo, LLC). All samples were gated on forward and side scatter, for singlets and for live cells.
8
Phenotyping Autologous CIK Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The phenotype of the autologous CIK cells was characterized by flow cytometry. CIK cells were resuspended at 2 × 105 cells per 100 μL of phosphate-buffered saline (PBS) and incubated for 30 min at 4°C with the following anti-human antibodies: anti-CD3-PE-Cy5, anti-CD4-FITC, anti-CD8-PE-CF594 and anti-CD56-PE-Cy7 (all from BD Bioscicence). The cells were analyzed using a CytomicsTM FC500 Flow Cytometer (Beckman Coulter, USA). Data analysis was performed with CXP analysis software (Beckman Coulter, USA).
9
Pneumococcal-specific CD4+ T cell Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Antigen-specific CD4+ T cell responses were measured as previously described.8 (link) In brief, non-adherent BAL cells (0.5x106cells/well) suspended in 200 µl of complete media were cultured in 96 well plates and stimulated with pneumococcal cell culture supernatant (8 µg/ml) (prepared from a standard encapsulated type 2 (D39) S. pneumoniae strain as previously described16 , 31 (link)). The cell culture supernatant has been previously utilised to probe pneumococcal-specific T cell immunity,8 (link), 15 (link), 16 , 31 (link) is rich in pneumococcal surface proteins and pneumolysin.31 (link) Cells were stained with Violet Viability dye (LIVE/DEAD® Fixable Dead Cell Stain kit, Invitrogen, UK), surface markers (anti-CD3 PE-Cy5, anti-CD4 APC-H7 and anti-CD8 PE-Cy7 antibodies, all from BD Bioscience, UK), and intracellular markers (anti-IFN-γ APC, anti-tumour necrosis factor (TNF) Alexa Fluor 488 and anti-IL17 PE antibodies, all from BD Bioscience, UK). Approximately 50,000 events were acquired in the CD4+ gate using a CyAn ADP 9-colour flow cytometer (Beckman Coulter, USA). Flow cytometry data were analysed using FlowJo software (TreeStar, USA).
10
Flow Cytometric Analysis of CIK Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
CIK cells were resuspended at 2 × 105 cells per 100 μL of phosphate-buffered saline (PBS) and incubated for 30 min at 4 °C with the following anti-human antibodies: anti-CD3-PE-Cy5, anti-CD4-FITC, anti-CD8-PE-CF594, anti-CD25-APC, anti-CD56-PE-Cy7, anti-CD45RO-APC, and anti-CD62L-FITC (all from BD Bioscicence). The cells were analyzed using a CytomicsTM FC500 Flow Cytometer (Beckman Coulter, USA). Data analysis was performed with CXP analysis software (Beckman Coulter, USA).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!