Trem2

Manufactured by Proteintech

Sourced in United States, China

TREM2 is a cell surface receptor expressed on myeloid cells, including microglia in the central nervous system. It plays a role in the regulation of inflammatory responses and phagocytic activity.

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Lab products found in correlation

Nx50 cryostat, by Thermo Fisher Scientific (1 mentions) Pan ck, by Novus Biologicals (1 mentions) Rneasy mini kit, by Qiagen (1 mentions) Odyssey fc system, by LI COR (1 mentions) β actin, by Merck Group (1 mentions) Rneasy ffpe kit, by Qiagen (1 mentions) Eclipse ti, by Nikon (1 mentions) F4 80, by Wuhan Servicebio Technology (1 mentions) Cd206, by Wuhan Servicebio Technology (1 mentions) Alexa fluor 488, by Wuhan Servicebio Technology (1 mentions) Kim 1, by Novus Biologicals (1 mentions)

3 protocols using trem2

Immunohistochemical Analysis of Tumor Samples

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Tumors were harvested and embedded in tissue molds containing OCT (Sakura) and frozen on dry ice prior to storage at −80°C. IHC staining were performed at CSHL Tissue Imaging Shared Facility. OCT embedded fresh tissue blocks were sectioned with Thermo #NX50 cryostat. 10μm thick sections were collected and mounted on positive charged glass slides (VWR superfrost plus micro slide) IHC slides were stained on DISCOVERY ULTRA IHC/ISH research platform (Roche) following standard protocols. Briefly, after fixation, slides were incubated with primary antibody at 37°C for 1h and Discovery multimer detection system (Discovery OmniMap HRP, Discovery DAB, Roche) was used to detect and amplify immuno-signals. Primary antibodies: Ki67 (Thermo Fisher 14-5698-82), 1:500 dilution; Pan-CK (Novus Bio NBP1-48348H), 1:100 dilution; TREM2 (Proteintech 13483-1-AP), 1:150 dilution.

Kleeman S.O., Thakir T.M., Demestichas B., Mourikis N., Loiero D., Ferrer M., Bankier S., Riazat-Kesh Y.J., Lee H., Chantzichristos D., Regan C., Preall J., Sinha S., Rosin N., Yipp B., de Almeida L.G., Biernaskie J., Dufour A., Tober-Lau P., Ruusalepp A., Bjorkegren J.L., Ralser M., Kurth F., Demichev V., Heywood T., Gao Q., Johannsson G., Koelzer V.H., Walker B.R., Meyer H.V, & Janowitz T. (2023). Cystatin C is glucocorticoid responsive, directs recruitment of Trem2+ macrophages, and predicts failure of cancer immunotherapy. Cell Genomics, 3(8), 100347.

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Quantitative Analysis of Liver and Macrophages

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Liver tissue was harvested and snap frozen at −80⁰C until further analysis. Bone marrow was extracted from femur and tibia and differentiated into bone marrow-derived macrophages (BMDMs) as described below and previously²⁵. For gene expression, total RNA was extracted from ~100 mg of liver tissue, from 1×10⁶ BMDMs or TG-Ms using RNeasy Mini Kits (Qiagen, Valencia CA, USA). Total RNA was also extracted from the formalin-fixed aortic arches used for en face analysis using an RNeasy FFPE Kit (Qiagen, Valencia, CA USA). 0.5–2 μg of RNA was reverse-transcribed, and the cDNA thus obtained was analyzed by real-time quantitative PCR by standard protocols using an ABI 7900HT instrument. For protein analyses, BMDMs were homogenized in cold RIPA buffer with protease inhibitors, SDS-PAGE was performed using equal amounts of protein, and Western blots were probed for Triggering receptor expressed on myeloid cells 2 (Trem2, Proteintech Group, Rosemont, IL, USA) and beta-actin (MilliporeSigma, St. Louis, MO, USA). Western blots were imaged using a LI-COR Odyssey Fc system and analyzed using Image Studio Software (Lincoln, NE, USA).

Chait A., Wang S., Goodspeed L., Gomes D., Turk K., Wietecha T., Tang J., Storey C., O’Brien K., Rubinow K., Tang C., Vaisar T., Gharib S., Lusis A, & den Hartigh L. (2021). Sexually dimorphic relationships among serum amyloid A3, inflammation, and cholesterol metabolism modulate atherosclerosis in mice. Arteriosclerosis, thrombosis, and vascular biology, 41(6), e299-e313.

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Immunofluorescence Analysis of Kidney Tissue

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For IF studies, kidney tissues were embedded at the optimal cutting temperature (OCT) immediately after the mice were euthanized. OCTembedded kidneys were cryosectioned into 5-μm sections and mounted on adhesion slides. The sections were fixed with cold methanol for 20 min, blocked with 5% v/v goat serum for 1 h at room temperature (RT), and incubated with primary antibodies against TREM2 (Proteintech, Wuhan, China), F4/80 (Servicebio, Wuhan, China), kidney injury molecule-1 (KIM-1; Novus, Port Orchard, WA, USA), inducible nitric oxide synthase (iNOS; Servicebio), CD206 (Servicebio), and Arg1 (Servicebio). The secondary antibodies used were Alexa Fluor 488 and Alexa Fluor 594 (Servicebio). The sections were then stained with 40, 6-diamidino-2phenylindole. All images were captured using a confocal microscope (ECLIPSE Ti; Nikon, Tokyo, Japan).

Cui Y., Chen C., Tang Z., Yuan W., Yue K., Cui P., Qiu X., Zhang H., Li T., Zhu X., Luo J., Sun S., Li Y., Feng C., Peng L., Xie X., Guo Y., Xie Y., Jiang X., Qi Z., Thomson A.W, & Dai H. (2024). TREM2 deficiency aggravates renal injury by promoting macrophage apoptosis and polarization via the JAK-STAT pathway in mice. Cell death & disease, 15(6).

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