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The NJTEN3 is a laboratory instrument designed for the measurement and analysis of various samples. It features a compact and durable design to support routine laboratory operations. The core function of the NJTEN3 is to provide accurate and reliable data for laboratory professionals.

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Lab products found in correlation

Mp6 xt22, by BioLegend (3 mentions) Ionomycin, by Merck Group (3 mentions) Phorbol myristate acetate (pma), by Merck Group (3 mentions) Cd80 16 10a1, by BioLegend (2 mentions) Cd45 30 f11, by BD (2 mentions) Cd86 gl 1, by BioLegend (2 mentions) Cd11c hl3, by BD (2 mentions) Cd8a 53 6, by Thermo Fisher Scientific (2 mentions) Cd274 10f 9g2, by BioLegend (2 mentions) M5 114, by BioLegend (2 mentions) Golgi stop plug, by BD (2 mentions) Cytofix cytoperm, by BD (2 mentions) Tc11 18h10, by BD (2 mentions) Xmg12, by BD (2 mentions) Cd4 rm4 5, by BioLegend (2 mentions) Facscanto 2 cytometer, by BD (2 mentions) Golgiplug, by BD (2 mentions) Intracellular staining kit, by BioLegend (1 mentions) Mp5 20f3, by BioLegend (1 mentions) Kaluza v1, by Beckman Coulter (1 mentions)

5 protocols using njten3

1

Pancreatic Lymph Node Cell Profiling

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Dissociated cells from pancreatic lymph nodes were suspended in buffer containing 2% FBS in PBS. To stain for surface antigens, cells were incubated with specific antibodies to F4/80 (BM-8, Biolegend), CD11c (HL3, BD Pharmigen), CD4 (RM4–5, Biolegend), CD45 (30- F11, BD Biosciences), CD8a (53–6.7, eBioscience), CD80 (16-10A1, Biolegend), CD274 (10F.9G2, Biolegend), CD86 (GL-1, Biolegend), and MHC-II (M5/114.15.2, Biolegend) or the appropriate isotype controls for 30 min. For the intracellular staining, cells were stimulated with 100ng/mL PMA (Sigma Aldrich), 500ng/ml Ionomycin (Sigma Aldrich), and Golgi Stop Plug (BD Pharmigen, 1/1000). After stimulation, cells were first stained for surface antigens, then fixed and permeabilized with BD Cytofix/Cytoperm™ (BD Pharmigen), according to the manufacturer’s recommendations. Cells were then incubated with specific antibodies to TNF-α (MP6-XT22, Biolegend), IL-1β (NJTEN3, Thermo Fisher Scientific), IL-17 (TC11-18H10, BD Pharmigen), IFN-γ (XMG12, BD Pharmigen), and FoxP3 (MF23, BD Pharmigen). After staining, the cells were washed, filtered, and analyzed on a FACS Canto II cytometer (BD). Data were analyzed using FlowJo software (Tree Star).
Piñeros A.R., Kulkarni A., Gao H., Orr K.S., Glenn L., Huang F., Liu Y., Gannon M., Syed F., Wu W., Anderson C.M., Evans-Molina C., McDuffie M., Nadler J.L., Morris M.A., Mirmira R.G, & Tersey S.A. (2022). Proinflammatory signaling in islet β cells propagates invasion of pathogenic immune cells in autoimmune diabetes. Cell reports, 39(13), 111011.
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2

Pancreatic Lymph Node Cell Profiling

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Dissociated cells from pancreatic lymph nodes were suspended in buffer containing 2% FBS in PBS. To stain for surface antigens, cells were incubated with specific antibodies to F4/80 (BM-8, Biolegend), CD11c (HL3, BD Pharmigen), CD4 (RM4–5, Biolegend), CD45 (30- F11, BD Biosciences), CD8a (53–6.7, eBioscience), CD80 (16-10A1, Biolegend), CD274 (10F.9G2, Biolegend), CD86 (GL-1, Biolegend), and MHC-II (M5/114.15.2, Biolegend) or the appropriate isotype controls for 30 min. For the intracellular staining, cells were stimulated with 100ng/mL PMA (Sigma Aldrich), 500ng/ml Ionomycin (Sigma Aldrich), and Golgi Stop Plug (BD Pharmigen, 1/1000). After stimulation, cells were first stained for surface antigens, then fixed and permeabilized with BD Cytofix/Cytoperm™ (BD Pharmigen), according to the manufacturer’s recommendations. Cells were then incubated with specific antibodies to TNF-α (MP6-XT22, Biolegend), IL-1β (NJTEN3, Thermo Fisher Scientific), IL-17 (TC11-18H10, BD Pharmigen), IFN-γ (XMG12, BD Pharmigen), and FoxP3 (MF23, BD Pharmigen). After staining, the cells were washed, filtered, and analyzed on a FACS Canto II cytometer (BD). Data were analyzed using FlowJo software (Tree Star).
Piñeros A.R., Kulkarni A., Gao H., Orr K.S., Glenn L., Huang F., Liu Y., Gannon M., Syed F., Wu W., Anderson C.M., Evans-Molina C., McDuffie M., Nadler J.L., Morris M.A., Mirmira R.G, & Tersey S.A. (2022). Proinflammatory signaling in islet β cells propagates invasion of pathogenic immune cells in autoimmune diabetes. Cell reports, 39(13), 111011.
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3

Assessing Endocytic and Cytokine Profiles of BMDMs

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After 2w with regular or high nitrate diet feeding, BMDMs were isolated and cultured as previously described [39] . Endocytic potential of BMDMs was assessed by uptake of fluorescently labeled dextran (ThermoFisher, Sweden). Briefly, BMDMs were cultured in 12-well plate for 24 h then incubated with Alexa Fluor 647-labeled Dextran (1 μg/ml) for 30 min. After washing away the extra dextran with PBS, the cells were detached with 2 mM EDTA. Cells were run in a Gallios flow cytometer (Beckman Coulter, Brea, CA) and analyzed using Kaluza v1.1 software (Beckman Coulter).
The intracellular cytokines of BMDMs was also measured by flow cytometer. Briefly, after 24 h of culture, the cells were first incubated with GolgiPlug (1 µl/ml, BD Biosciences) in complete DMEM for 4 h at 37 °C before incubation with antibodies. Fixation/Permeablization and intracellular staining was conducted using the eBioscience Intracellular Staining Kit and the following antibodies: IL-6 (MP5-20F3, Biolegend), IL-1 beta pro-form (NJTEN3, eBioscience), and TNF alpha (MP6-XT22, Biolegend).
Yang T., Zhang X.M., Tarnawski L., Peleli M., Zhuge Z., Terrando N., Harris R.A., Olofsson P.S., Larsson E., Persson A.E., Lundberg J.O., Weitzberg E, & Carlstrom M. (2017). Dietary nitrate attenuates renal ischemia-reperfusion injuries by modulation of immune responses and reduction of oxidative stress. Redox Biology, 13, 320-330.
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4

Immunophenotyping of Immune Cells

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Antibodies used were against 5-lipoxygenase (Cayman Chemicals, catalog number 160402), autotaxin (Phoenix Pharmaceuticals, catalog number H-008-29), CD45 (Miltenyi Biotech, clone 30F11.1 catalog number 130-091-609), CD80 (Biolegend, clone 16-10A1 catalog number 104706), CD86 (Biolegend, clone GL-1 catalog number 105007), CD206 BioRad, clone MR5D3 catalog number MCA2235GA, and Biolegend, clone C068C2 catalog number 141707), F4-80 (Biolegend clone BM8 catalog number 123107), IL1β (eBiosciences, clone NJTEN3 catalog number) 11-7114-82, Ly6C (eBiosciences, clone HK1.4 catalog number 17-5932-80), Ly6G (BioLegend, clone 1A8 catalog number 127613), and TNFα (Miltenyi Biotech, clone REA636 catalog number 130-109-720).
Kern K., Schäfer S.M., Cohnen J., Pierre S., Osthues T., Tarighi N., Hohmann S., Ferreiros N., Brüne B., Weigert A., Geisslinger G., Sisignano M, & Scholich K. (2018). The G2A Receptor Controls Polarization of Macrophage by Determining Their Localization Within the Inflamed Tissue. Frontiers in Immunology, 9, 2261.
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5

Multiparametric Flow Cytometry Analysis

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A single-cell suspension was prepared from the spleen and red blood cells were lysed in PharmLyse (Becton Dickinson, Franklin Lakes, NJ). Fc receptors were blocked with an anti-CD16/CD32 (Becton Dickinson) antibody and the cells were then stained with anti-mouse antibodies against Gr-1 (clone: RB6-8C5) (eBioscience, San Diego, CA), Ly6G (1A8), Ly6C (AL-21), CD11b (M1/70), CD3 (17A2), CD4 (RM4-5), CD25 (PC61), and Foxp3 (MF23) (Becton Dickinson). Dead cells (stained positive by propidium iodide) were excluded from the analysis. To examine intracellular cytokine staining, 1 × 10 6 splenocytes were stimulated with phorbol myristate acetate (5 ng/mL)/ionomycin (500 ng/mL) (Sigma-Aldrich, St. Louis, MO) in the presence of GolgiPlug (Becton Dickinson) and then cultured in RPMI 1640 (Wako, Osaka, Japan) for 4 h at 37°C. Subsequently, cells were stained with antibodies against intracellular IL-1β (NJTEN3) (eBiosciences), IL-2 (JES6-5H4), IL-4 (11B11), IL-6 (MP5-20F3), IFNγ (XMG1.2), and TNFα (MP6-XT22) (Becton Dickinson). Rat IgG1κ and 2bκ (Becton Dickinson) were used as negative controls. Samples were run in a Calibur flow cytometer (Becton Dickinson) and data were analyzed using FlowJo software (TreeStar, Inc., Ashland, OR).
Nakamura T., Nakao T., Yoshimura N, & Ashihara E. (2015). Rapamycin Prolongs Cardiac Allograft Survival in a Mouse Model by Inducing Myeloid-Derived Suppressor Cells. American journal of transplantation : official journal of the American Society of Transplantation and the American Society of Transplant Surgeons, 15(9).
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