Cobas 6000 with ise module

Manufactured by Roche

The Cobas 6000 with ISE module is an automated clinical chemistry and immunoassay analyzer designed for high-throughput laboratory testing. The core function of the Cobas 6000 is to perform a variety of clinical chemistry and immunoassay tests on patient samples. The ISE module specifically handles the analysis of electrolytes, such as sodium, potassium, and chloride.

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Lab products found in correlation

Triton x 100, by Merck Group (1 mentions) Cytotoxicity detection kit, by Roche (1 mentions) Yo pro 1 iodide, by Thermo Fisher Scientific (1 mentions) Yo pro 1, by Agilent Technologies (1 mentions) Nitrocellulose membrane, by Merck Group (1 mentions) Bis tris gradient gel, by Thermo Fisher Scientific (1 mentions)

Spelling variants of this product

Cobas 6000 (699 citations) ISE module (5 citations) Roche module Cobas 6000 (2 citations)

2 protocols using cobas 6000 with ise module

Quantifying Cell Membrane Integrity

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The presence of lactate dehydrogenase (LDH) in cell culture supernatants was measured using the Cytotoxicity Detection kit (Roche), following manufacturer’s instructions. It was expressed as the percentage of the total amount of LDH present in the cells. For Yo-Pro uptake, macrophages were preincubated for 5 min at 37 °C with 2.5 µM of Yo-Pro-1 iodide (Life Technologies) after galvanic current application or 1% triton X100 (Sigma-Aldrich) application. Yo-Pro-1 fluorescence was measured after the treatments every 5 min for the first 30 min and then every 30 min for the following 3 h with an excitation wavelength of 478 ± 20 nm and emission of 519 ± 20 nm in a Synergy neo2 multi-mode plate reader (BioTek). Intracellular K⁺ was quantified using macrophages lysates, as has already been reported (Compan et al., 2012 (link)), and measured by indirect potentiometry on a Cobas 6000 with ISE module (Roche).

Peñin-Franch A., García-Vidal J.A., Martínez C.M., Escolar-Reina P., Martínez-Ojeda R.M., Gómez A.I., Bueno J.M., Minaya-Muñoz F., Valera-Garrido F., Medina-Mirapeix F, & Pelegrín P. (2022). Galvanic current activates the NLRP3 inflammasome to promote Type I collagen production in tendon. eLife, 11, e73675.

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Co-immunoprecipitation and ion quantification

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Detailed methods used for Western blot analysis, caspase-1 activity and ELISAs have been described previously (26 (link), 27 (link)). For coIP, equal amounts of protein (500–750 μg) were incubated with 2 μl (1.5 μg) of anti-GFP (Calbiochem) for 1 h at 4ºC on a rotating wheel. After adding prot-G-agarose beads (20 μl per sample) for 1 h at 4ºC, beads were washed three times (5 min at 4ºC) in lysis buffer. Proteins were eluted by heating samples (5 min at 80ºC) in presence of reducing agent and loading sample buffer (Invitrogen). Proteins were separated on 4-12% gradient Bis-tris gels (Invitrogen) and transferred to a nitrocellulose membrane (Millipore). Intracellular K⁺ was quantified from THP-1 cell lysates by indirect potenciometry on a Cobas 6000 with ISE module (Roche).

Compan V., Martín-Sánchez F., Baroja-Mazo A., López-Castejón G., Gomez A.I., Verkhratsky A., Brough D, & Pelegrín P. (2014). Apoptosis-associated speck-like protein containing CARD forms specks but does not activate caspase-1 in the absence of NLRP3 during macrophage swelling. Journal of immunology (Baltimore, Md. : 1950), 194(3), 1261-1273.

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