Crid3

Manufactured by Merck Group

Sourced in United States

CRID3 is a laboratory equipment product offered by Merck Group. It is designed for the purpose of conducting research and analysis. The core function of CRID3 is to provide a controlled and consistent environment for various experimental procedures.

Automatically generated - may contain errors

Lab products found in correlation

Z vad, by Merck Group (1 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions) Lps escherichia coli 0111 b4, by InvivoGen (1 mentions) Caspase 1 antibody, by Adipogen (1 mentions)

2 protocols using crid3

Bovine Macrophage Infection Assays

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The bovine macrophages were cultured on 24-well plates, at a density of 3 × 10⁵ cells per well, to perform infection assays. We used multiplicities of infection (MOIs) of 2:1 and of 10:1 for the NY-1 ncp-BVDV strain and 2:1 for NADL cp-BVDV strain, with 500µL of RPMI medium supplemented with 10% heat-inactivated FBS (Gibco, New York, NY, USA). The negative controls were macrophages cultured with RPMI and 10% FBS, and the positive control was treated with 300 ng/mL of LPS from E. coli type O26:B26. For the inhibition treatments, the macrophages were pre-incubated for 2 h with CRID3 (Cytokine Release Inhibitory Drug 3, 50 µM; Sigma Aldrich, St. Louis, MO, USA), Y-VAD (50 µM; Merck, Boston, MA, USA) or Z-VAD (carbobenzoxy-valyl-alanyl-aspartyl-[O-methyl]- fluoromethylketone, 50 µM; Sigma Aldrich, St. Louis, MO, USA) and ODN A151 (5′-TTAGGGTTAGGGTTAGGGTTAGGG-3′) (3 µM, 6 µM and 12 µM). The supernatants were recovered at 24 h post-infection.

Gallegos-Rodarte C., Escobar-Chavarría O., Cantera-Bravo M.M., Sarmiento-Silva R.E, & Benitez-Guzman A. (2023). NLRP3 Inflammasome Involved with Viral Replication in Cytopathic NADL BVDV Infection and IFI16 Inflammasome Connected with IL-1β Release in Non-Cytopathic NY-1 BVDV Infection in Bovine Macrophages. Viruses, 15(7), 1494.

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Microglial Activation and Tau/Aβ Responses

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For IL-1β analysis, 75,000 microglia/well in 96-well plate were used. For Caspase-1 detection or collection of conditioned medium for neuronal experiments, 2 million microglia/well were plated in a 6-well plate. Microglia were activated with 100ng/ml ultrapure LPS (Escherichia coli 0111:B4, Invivogen) for 3 hours. After washing with warm DPBS, recombinant tau monomers, oligomers or fibrils at a concentration of 2μM or 10μM Aβ fibrils were added for 6 hours to the cultures before collection of conditioned media. For CRID3 (SigmaAldrich) treatments, the inhibitor was mixed in with the tau treatments at a concentration of 100nM. Conditioned media were analyzed for IL-1β secretion by ELISA (R&D Systems) according to the manufacturer’s instructions or for Caspase-1 secretion by using a Jess system (ProteinSimple). Here, samples were run undiluted on a 12–230 kDa separation module according to the manufacturer’s instructions and detection was achieved by using a Caspase-1 antibody (Adipogen) at 1:50. Data were analyzed with Compass software version 4.0.0 (ProteinSimple).

Ising C., Venegas C., Zhang S., Scheiblich H., Schmidt S.V., Vieira-Saecker A., Schwartz S., Albasset S., McManus R.M., Tejera D., Griep A., Santarelli F., Brosseron F., Opitz S., Stunden J., Merten M., Kayed R., Golenbock D.T., Blum D., Latz E., Buée L, & Heneka M.T. (2019). NLRP3 inflammasome activation drives tau pathology. Nature, 575(7784), 669-673.

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