Nur77

Manufactured by Thermo Fisher Scientific

The Nur77 is a laboratory instrument designed to measure and analyze Nur77 expression levels in biological samples. It utilizes advanced detection technologies to quantify the presence and activity of this important molecular target. The Nur77 provides researchers with reliable and precise data to support their investigations into cellular signaling pathways and transcriptional regulation.

Automatically generated - may contain errors

Lab products found in correlation

6 protocols using nur77

Murine CD8+ T Cell Immunophenotyping

Check if the same lab product or an alternative is used in the 5 most similar protocols

The following antibodies and corresponding isotype controls were used in this study: CD8α (53–6.7 Tonbo), CD44 (1M7; BioLegend), CD45.2 (104; Tonbo), CD103 (2E7; BioLegend), CD69 (H1.2F3; BioLegend), IFN-γ (XMG1.2; Tonbo), Thy1.1 (OX-7; BioLegend), Thy1.2 (53–2.1; BioLegend), CD127 (A7R34; BioLegend), KLRG1 (2F1; Tonbo), Nur77 (12.14; eBioscience), CD3ε (17A2; BioLegend), Ly6C (HK1.4; BioLegend), Ly6G (IA8; BioLegend), and CD11b (M1/70; BioLegend). H-2D^b-GP_33-41 and H2-K^b-OVA_257-264 tetramers were provided by J. Harty (University of Iowa, Iowa City, IA). H-2K^b-B8R_20-27 tetramer was obtained from the National Institutes of Health tetramer core facility. Staining for surface antigens was performed in PBS/1% fetal bovine serum for 15 min at 4°C. For tetramer binding, cells were incubated for 45 min at room temperature Data were acquired using either a Fortessa or LSRII Flow Cytometer (BD) in the OHSU Flow Cytometry Core Facility. Flow cytometry data were analyzed using FlowJo software, version 9.9. For depletion of Thy1.1-expressing CD8⁺ T cells, mice were treated with 2–3 µg of control rat IgG (Sigma-Aldrich) or anti-Thy1.1 antibody (clone 19E12; BioXCell) in 200 µl of PBS by intraperitoneal injection.

Khan T.N., Mooster J.L., Kilgore A.M., Osborn J.F, & Nolz J.C. (2016). Local antigen in nonlymphoid tissue promotes resident memory CD8+ T cell formation during viral infection. The Journal of Experimental Medicine, 213(6), 951-966.

+ Open protocol

+ Expand

Comprehensive Immune Cell Profiling

Check if the same lab product or an alternative is used in the 5 most similar protocols

Fluorochrome-conjugated antibodies against CD3, CD4, CD5, CD8a, CD27, CD45RA, CD45RO, CD122, Nur77, Eomes, NKG2A, NKG2D, KIR2D and KIR3DL1 were purchased from eBioscience, Biolegend or Miltenyi Biotec.

White J.T., Cross E.W., Burchill M.A., Danhorn T., McCarter M.D., Rosen H.R., O'Connor B, & Kedl R.M. (2016). Virtual memory T cells develop and mediate bystander protective immunity in an IL-15-dependent manner. Nature Communications, 7, 11291.

+ Open protocol

+ Expand

Isolation and Characterization of Murine Microglia

Check if the same lab product or an alternative is used in the 5 most similar protocols

Microglia were isolated as previously described (21 (link)). Anesthetized mice were perfused intracardially with PBS. Brain and spinal cord were homogenized and isolated cells were separated using a density gradient. After intense washing steps, cells were incubated with Fc Block [TruStain fcX 9 (93); BioLegend, London, U.K.] at 4°C for 10 min, followed by surface staining in PBS containing 5% FCS for 30 min at 4°C. Intracellular staining of Nur77 was performed by using a Foxp3/transcription factor staining buffer set (eBioscience, San Diego, CA). Cells were stained according to the manufacturer’s instructions. Microglia were stained with the indicated FACS Abs: Nur77 (12.14, eBioscience), CD39 (Duha59), CD206 (C068C2), Ly6C (HK1.4), Ly6G (1A8), CD86 (2331), MHC class II (L243), CD11b (M1/70), and CD45 (30-F11) (all BioLegend), as well as Live/Dead (Life Technologies). For in vitro experiments, isolated cells were cultured for 3 wk in appropriate DMEM/F12 medium containing 10% FCS and M-CSF to obtain microglia. Following treatment with either 1 mM of ATP (A6419; Sigma-Aldrich) and/or 100 ng/ml LPS (L-2630; Sigma-Aldrich), microglia were further analyzed.

Rothe T., Ipseiz N., Faas M., Lang S., Perez-Branguli F., Metzger D., Ichinose H., Winner B., Schett G, & Krönke G. (2017). The Nuclear Receptor Nr4a1 Acts as a Microglia Rheostat and Serves as a Therapeutic Target in Autoimmune-Driven Central Nervous System Inflammation. Journal of immunology (Baltimore, Md. : 1950), 198(10), 3878-3885.

+ Open protocol

+ Expand

Multiparametric Flow Cytometry of Immune Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

Single-cell suspensions of thymus, spleen, or lymph nodes were prepared and red blood cells were lysed. Cells were incubated with various combinations of the following antibodies: CD3, CD4, CD5, CD8, CD19, FoxP3, Nur77, IFN-γ (eBioscience, San Diego, Calif, and Tonbo, San Diego, Calif). Samples were collected using FACS LSRII (BD, San Jose, Calif) or sorted using FACSAria (BD), and data were analyzed with FlowJo software (Tree Star, Ashland, Ore). Differences between groups were analyzed using 2-tailed t test (Graphpad Prism software, La Jolla, Calif).

Ku M., Ke E., Sabouri-Ghomi M., Abadejos J.R., Freeman B., Nham A., Phillips N., Yang K.Y., Lui K.O, & Kirak O. (2018). Deconstructive somatic cell nuclear transfer reveals novel regulatory T-cell subsets. The Journal of allergy and clinical immunology, 142(3), 997-1000.e4.

+ Open protocol

+ Expand

Activation-Induced Nur77 Expression

Check if the same lab product or an alternative is used in the 5 most similar protocols

Murine thymocytes and Jurkat cells were stimulated with PMA (20ng mL⁻¹) and ionomycin (1µM) or C305, an anti-Vβ8 mouse mAb (IgM) that reacts with the Jurkat TCR β chain (21 (link)) for two hours at 37°C. Cells were lysed directly in SDS-PAGE sample buffer after stimulation. Proteins were separated on SDS-PAGE gels, transferred to Immobilon-P polyvinylidene difluoride membranes (Millipore) by standard immunoblotting techniques. Primary staining was performed with the following antibodies: Nur77 (eBioscience) and GAPDH (Santa Cruz). Horseradish peroxidase (HRP)-conjugated secondary antibodies were from Southern Biotech and visualized with SuperSignal ECL reagent or SuperSignal West Femto maximum sensitivity substrate (Pierce Biotechnology) on Chemi-Doc Image Lab station (Bio-Rad).

Ashouri J.F, & Weiss A. (2016). Endogenous Nur77 is a specific indicator of antigen receptor signaling in human T and B cells. Journal of immunology (Baltimore, Md. : 1950), 198(2), 657-668.

+ Open protocol

+ Expand

Thymocytes TCR Activation Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols

Thymocytes isolated from miR-181a^+/− and miR-181a^−/− mice were left untreated or were stimulated for 3 h at 37 °C in the presence of plate-bound αCD3 (17A2, 2.5 μg/ml) and soluble αCD28 (37.51, 2.5 μg/ml). Next, cells were stained for surface CD4 and CD8α and intracellular Nur77 with intracellular staining buffer set (eBioscience).

Łyszkiewicz M., Winter S.J., Witzlau K., Föhse L., Brownlie R., Puchałka J., Verheyden N.A., Kunze-Schumacher H., Imelmann E., Blume J., Raha S., Sekiya T., Yoshimura A., Frueh J.T., Ullrich E., Huehn J., Weiss S., Gutierrez M.G., Prinz I., Zamoyska R., Ziętara N, & Krueger A. (2019). miR-181a/b-1 controls thymic selection of Treg cells and tunes their suppressive capacity. PLoS Biology, 17(3), e2006716.

+ Open protocol

+ Expand

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?

Revolutionizing how scientists
search and build protocols!