γ xmg1.2

Manufactured by Thermo Fisher Scientific

The γ (XMG1.2) is a compact handheld gamma radiation detector designed for general radiation monitoring applications. It features a sodium iodide (NaI) scintillation detector and provides measurements of gamma radiation levels. The device offers basic functionality for radiation detection without further interpretation or extrapolation on its intended use.

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Lab products found in correlation

Ebio17b7, by Thermo Fisher Scientific (2 mentions) Foxp3 staining kit, by Thermo Fisher Scientific (2 mentions) Ionomycin, by Merck Group (2 mentions) Aqua live dead stain, by Thermo Fisher Scientific (1 mentions) Lsrii analyzer, by BD (1 mentions) Perm wash buffer, by BD (1 mentions) Fluorescein isothiocyanate (fitc), by Thermo Fisher Scientific (1 mentions) Pe conjugated antibodies specific, by BD (1 mentions) Rm4 5, by Thermo Fisher Scientific (1 mentions) Anti actin c4, by Merck Group (1 mentions) Celltrace cfse cell proliferation kit, by Thermo Fisher Scientific (1 mentions) Interleukin 6 (il 6), by R&D Systems (1 mentions) Mg132, by Cayman Chemical (1 mentions) Interleukin 2 (il 2), by R&D Systems (1 mentions) Relb cflag pcdna3, by Addgene (1 mentions) P52 cflag pcdna3, by Addgene (1 mentions) C rel cflag pcdna3, by Addgene (1 mentions) Hsp60 h 1, by Santa Cruz Biotechnology (1 mentions) Monensin, by Merck Group (1 mentions) Jes6 5h4, by Thermo Fisher Scientific (1 mentions)

6 protocols using γ xmg1.2

Comparative Immune Profiling of Mice

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Spleens, lymph nodes (inguinal plus axillary), and thymi were harvested from age-matched female IL-11Rα^+/+ and IL-11Rα^−/− mice and were dissociated into a single cell suspension. CNS mononuclear cells were isolated from spinal cords and cerebellums (pooled from multiple mice within each group) using collagenase digestion followed by Percoll gradient [29] (link). These cells were stained using the aqua live/dead stain (Invitrogen, Burlington, ON) and antibodies specific for the following cell surface markers (all from eBioscience): CD4 (GK1.5), CD8 (53–6.7), B220 (RA3-6B2), CD11c (N418), CD62L (MEL-14), CD44-PE (IM7) using standard protocols provided by eBioscience. CD4⁺CD25⁺FoxP3⁺ Treg were stained using reagents and antibodies provided with the FoxP3-staining kit (eBioscience) along with an antibody specific for ICOS (7E.17G9). Intracellular staining for IL-17A (eBio17B7) and IFN-γ (XMG1.2) was performed using antibodies from eBioscience and Perm/Wash buffer and staining protocols from BD Pharmingen. Data were acquired using the LSRII analyzer (BD Biosciences, Mississauga, ON) and were analyzed with FlowJo software (Treestar, Ashland, OR). Gates were set using fluorescence minus one controls.

Figueiredo C.A., Drohomyrecky P.C., McCarthy S.D., Leontyev D., Ma X.Z., Branch D.R, & Dunn S.E. (2014). Optimal Attenuation of Experimental Autoimmune Encephalomyelitis by Intravenous Immunoglobulin Requires an Intact Interleukin-11 Receptor. PLoS ONE, 9(7), e101947.

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ASK1 Pathway Regulation in HEK293 and BMDMs

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HEK293 cells were cultured in Dulbecco's modified Eagle's medium (DMEM) containing 10% fetal bovine serum (FBS), 4.5 mg/ml glucose and 100 units/ml penicillin G. A crude population of BMDMs was generated in vitro from mouse bone marrow as described30 (link). PE-conjugated antibodies specific for MHC class II (M5/114.15.2), B220 (RA3-6B2) and IL-17 (TC11-18H10) were purchased from BD PharMingen. Phycoerythrin (PE)-Cy5-, PE-, and FITC-conjugated antibodies against CD4 (GK1.5) and IFN-γ (XMG1.2) were purchased from eBioscience. Phospho-ASK1 antibody that recognizes the activating phosphorylation of ASK1 was described previously19 (link). DNFB and DNBS were purchased from Sigma-Aldrich. FITC was purchased from Thermo Scientific. 4-Amino-1-tert-butyl-3-(1′-naphthyl)pyrazolo[3,4-d]pyrimidine (1Na-PP1) and 4-amino-1-tert-butyl-3-(1′-naphthylmethyl)pyrazolo[3,4-d]pyrimidine (1NM-PP1) were purchased from Calbiochem. The ASK1 inhibitor K811 was provided by Kyowa Hakko Kirin Co., Ltd20 (link).

Mizukami J., Sato T., Camps M., Ji H., Rueckle T., Swinnen D., Tsuboi R., Takeda K, & Ichijo H. (2014). ASK1 promotes the contact hypersensitivity response through IL-17 production. Scientific Reports, 4, 4714.

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Antibody-based Immune Cell Analysis

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Functional-grade antibodies for CD3 (145-2C11), CD28 (37.51) and IFN-γ (XMG1.2) were from eBioscience, and the anti-IL-4 antibody (11B11) was from National Cancer Institute Preclinical Repository. Fluorescence-labeled antibodies for CD3 (17A2), CD4 (RM4-5), CD8 (53-6.7), CD11b (M1/70), CD44 (IM7), CD45 (30-F11), B220 (RA3-6B2), CD62L (MEL-14), CD69 (H1.2F3), TCRβ (H57-597), Foxp3 (FJK-16s), IL-17A (eBio17B7), and IFN-γ (XMG1.2) were purchased from eBioscience. Antibodies for mouse NIK (H248, 1:1,000) and Tubulin (TU-02, 1:2000) were from Santa Cruz Biotechnology. Anti-Actin (C-4, 1:10,000) was from Sigma, and anti-p100/p52 (TB4, 1:8,000) was from National Cancer Institute Preclinical Repository. CellTrace™ CFSE Cell Proliferation Kit was from Thermo Fisher Scientific. The recombinant mouse cytokines IL-2, IL-4, IL-6, IL-12, and TGF-β were from R&D systems. MG132 was from Cayman Chemical.

Li Y., Wang H., Zhou X., Xie X., Chen X., Jie Z., Zou Q., Hu H., Zhu L., Cheng X., Brightbill H.D., Wu L.C., Wang L, & Sun S.C. (2016). Cell intrinsic role of NF-κB-inducing kinase in regulating T cell-mediated immune and autoimmune responses. Scientific Reports, 6, 22115.

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Generating NF-κB Signaling Plasmids and Antibodies

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pMIGR1-mP52 and pMIGR1-mGM-CSF were generated by inserting mouse p52 (encoding amino acids 1–405 of NF-κB2) and mouse Csf2 cDNAs, respectively, into the EcoRI and BglII sites of pMIGR1-GFP vector. pcDNA-based expression vectors encoding FLAG-tagged mouse p52 (p52-cFlag-pcDNA3), mouse c-Rel (c-Rel-cFlag-pcDNA3), and mouse RelB (RelB-cFlag-pcDNA3) were from Addgene. The Csf2 promoter luciferase plasmid (pGL3-mCsf2), covering the region −225 to +26 of mouse Csf2 promoter (15 (link)), was provided by Dr. Takeshi Matsumura (Kumamoto University).
Functional grade anti–mouse (m) CD3ɛ (145-2C11) and anti-mCD28 (37.51) antibodies and blocking antibodies for mIFN-γ (XMG1.2) and mIL-4 (11B11) were from eBioscience. Fluorescence-labeled antibodies for mCD4 (L3T4), mCD8 (53-6.7), mCD3 (145-2C11), CD44 (IM7), CD62L (MEL-14), IL-17A (eBio17B7), GM-CSF (MP1-22E9), and IFN-γ (XMG1.2) were purchased from eBioscience. Antibodies for mouse p50 (C-19), c-Rel (sc-70), RelB (C-19), NIK (H-248), Lamin B (C-20), and Hsp60 (H-1) were from Santa Cruz Biotechnology. An antibody recognizing both p100 and p52 (Anti-p52/p100) was provided by NCI Preclinical Repository.

Yu J., Zhou X., Nakaya M., Jin W., Cheng X, & Sun S.C. (2014). T Cell Intrinsic Function of the Noncanonical NF-κB Pathway in the Regulation of GM-CSF Expression and EAE Pathogenesis. Journal of immunology (Baltimore, Md. : 1950), 193(1), 422-430.

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Phenotypic Analysis of Dendritic Cells

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For phenotypic analysis of DCs, splenic lymphocyte suspensions were stained with fluorochrome conjugated antibodies specific for the following surface markers: CD11c (N418), CD80 (16–10 A1), CD86 (GL1), CD40 (39164), MHCII (10.2.16), and PDL1 (10F.9G2), all of which were purchased from eBioscience (eBioscience, San Diego, CA, USA). Cell suspensions of thymus, spleen, and pancreatic lymph nodes (PLNs) were stained and analyzed with antibodies against CD4 (RM4-5), CD8α (53-6.7), CD25 (PC61.5), Foxp3 (FJK-16s), IFN-γ (XMG1.2), IL-4 (11B11), IL-2 (JES6-5H4), programmed cell death protein-1 (PD-1) (J43), T-bet (4B10), Eomesodermin (Eomes) (Dan11mag), CD160 (CNX46-3), CD244 (C9.1), CD272 (8F4), KLRG1, or killer cell lectin-like receptor G1 (2F1), all purchased from eBioscience. Antibodies against TNF-α (MP6-XT22), LAG3 or lymphocyte-activation gene 3 (C9B7W), and TIM3 or T cell immunoglobulin and mucin domain 3 (RMT3-23) were purchased from BioLegend (San Diego, CA, USA). For intracellular cytokine staining, splenocytes were stimulated with phorbol 12-myristate 13-acetate/PMA, ionomycin, and monensin (Sigma-Aldrich, St. Louis, MO, USA) for 6 h, followed by staining with antibodies. All analyses were performed on a FACSCalibur flow cytometer (BD Biosciences, San Jose, CA, USA) and analyzed using CellQuest software (BD Biosciences).

Kulshrestha D., Yeh L.T., Chien M.W., Chou F.C, & Sytwu H.K. (2017). Peripheral Autoimmune Regulator Induces Exhaustion of CD4+ and CD8+ Effector T Cells to Attenuate Autoimmune Diabetes in Non-Obese Diabetic Mice. Frontiers in Immunology, 8, 1128.

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Comprehensive Immune Cell Profiling

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Total CNS infiltrates, spleen, and LN cells were washed once in FACS buffer (PBS containing 0.1% BSA) before Fc receptor blocking with anti-CD16/CD32 (93; eBioscience, or 2.4G2; BD PharMingen). Staining of surface molecules was performed at room temperature using antibodies against CD4 (RM4-5), CD8α (53-6.7), CD90.1 (OX-7), α4β7 (LPAM-1, DATK32), CD25 (PC61), CD62L (MEL-14), CD69 (H1.2F3), CD45 (30-F11), CD11b (M1/70), PD1 (J43), CD27 (LG.3A10), and TIGIT (Vstm3; differently conjugated; all eBioscience, BioLegend, or BD) in FACS buffer for 15 min in the dark. Intracellular Foxp3 (FJK-16s, PE- or APC-conjugated; eBioscience) staining was performed using the Foxp3 staining kit (eBioscience) according to the manufacturer’s instructions. Intracellular cytokine staining of splenocytes with APC-conjugated IL-2 (JES6-5H4), IFN-γ (XMG1.2), and IL-10 (JES5-16E3) and PE-conjugated IL-4 (11B11), IL-17 (eBio17B7), GM-CSF (MP1-22E9), and TNFα (MP6-XT22; all eBioscience) was performed after 3–6-h restimulation with T/I (tetradecanoyl phorbol acetate [10 ng/ml; Sigma-Aldrich] plus ionomycin [5 nM; Merck Biosciences]) in the presence of GolgiStop and GolgiPlug (both BD Pharmingen) using the IC Fixation Buffer Kit (eBioscience). Data were acquired on a FACS Canto II (BD Biosciences) flow cytometer and analyzed with FlowJo software (TreeStar).

Xiao Y., Qureischi M., Dietz L., Vaeth M., Vallabhapurapu S.D., Klein-Hessling S., Klein M., Liang C., König A., Serfling E., Mottok A., Bopp T., Rosenwald A., Buttmann M., Berberich I., Beilhack A, & Berberich-Siebelt F. (2020). Lack of NFATc1 SUMOylation prevents autoimmunity and alloreactivity. The Journal of Experimental Medicine, 218(1), e20181853.

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