Examinations of serum antibodies were used by each physician to evaluate the past infection condition and/or the possibility of vaccination. Serum samples obtained at initial visits were used for measurement of anti-SARS-CoV-2 antibody using the Elecsys Anti-SARS-CoV-2 S (S300) electrochemiluminescence (ECLIA) kit (Roche Diagnostics, Rotkreuz, Switzerland) and cobas 8000 modular analyzer system at the Central Laboratory of Okayama University Hospital. The measurement range is from 0.40 U/mL to 25,000 U/mL (with 1:100 dilution) with a concentration of <0.80 U/mL considered negative. Regarding the unit exchanges of the antibody titer, since the unit value derived from this ECLIA kit has the highest linearity range to the binding antibody unit (BAU) based on the WHO International Standard [23 (link)] for COVID-19 serological tests (Elecsys IU = 0.972 × BAU: R = 0.9996), the present unit “U/mL” substantially represents “BAU/mL”. The thresholds of the antibody titers were set to 50, 100, 200, and 500 U/mL to stratify the patients based on a past report [24 (link)].
Cobas 8000 modular analyzer system
Manufactured by Roche
Sourced in Switzerland, Germany
The Cobas 8000 modular analyzer system is a comprehensive in-vitro diagnostic solution designed for high-volume clinical laboratories. It is capable of performing a wide range of clinical chemistry and immunochemistry tests. The system features a modular design, allowing for customization to meet the specific needs of individual laboratories.
Automatically generated - may contain errors
6 protocols using cobas 8000 modular analyzer system
1
Quantifying Anti-SARS-CoV-2 Antibodies
2
BaCl2 Injection: Serum Biomarker Profiling
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At day 0, as a control group before BaCl2 injection, and at days 1 and 7 after BaCl2 injection, whole blood was collected in serum-separating tubes by cardiac puncture. From the separated serum after centrifugation, levels of chemical markers including aspartate aminotransferase (AST), alanine aminotransferase (ALT), total cholesterol, creatinine kinase (CK), creatinine, glucose, C-reactive protein (CRP), and lactate dehydrogenase (LDH) were measured using a Cobas 8000 modular analyzer system (Roche, Basel, Switzerland) according to the manufacturer’s protocol.
3
Serum and Ascites Biomarker Analysis
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Levels of C-reactive protein (CRP), albumin (ALB), triglycerides (TRIG), cholesterol (CHOL), low-density lipoprotein (LDL), and high-density lipoprotein (HDL) were assessed in serum and cell free ascites supernatants. Frozen samples (−80°C) were thawed, diluted with 1x PBS (1:5 for plasma, 1:2 for ascites supernatants) and analyzed on a cobas 8000 modular analyzer system (Hoffmann-LaRoche, Basel, Switzerland) using the respective tests, distributed by Roche diagnostics (CRPL3, ALBT2, TRIGL, CHOL2, LDL_C, and HDLC3) in an ISO9001:2008 certified laboratory (Department of Laboratory Medicine, Medical University of Vienna).
4
Renal Function and Histology Assessment
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In mouse serum, blood urea nitrogen (BUN) and creatinine (Cr) levels were evaluated by GCLabs (Yongin, Korea) using the Cobas 8000 modular analyzer system (Roche, Germany). Kidney tissues from each experimental group were immersion-fixed with 4% paraformaldehyde (pH 7.4) and then embedded in paraffin. Two-micrometer tissue sections were prepared and stained with periodic acid-Schiff (PAS) and Masson’s trichrome using standard protocols for the determination of histological changes and collagen deposition, respectively. Immunohistochemical analysis of kidney tissues detected the Nox-1 (1:100, ab121009, Abcam) and Nox-4 proteins (1:100, MA5-32090, Invitrogen).
5
Serum Biomarker Analysis in Mice
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At the end of the experimental period, blood samples from each mouse were collected into tubes by cardiac puncture. The blood was sampled into ethylenediaminetetraacetic acid-free bottles for serum separation. The blood urea nitrogen (BUN), creatinine (Cr), uric acid, total cholesterol (TC), low-density lipoprotein (LDL) and triglycerides level in the serum were measured by GCLabs (Yongin, Korea) using the Cobas 8000 modular analyzer system (Roche, Basel, Switzerland).
6
Kidney Histological and Functional Analysis
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All mice were euthanized by cardiac puncture under anesthesia 7 weeks after reperfusion. Blood samples and the kidneys were collected for analysis. After centrifuging each blood sample, the serum underwent biochemical analysis. Blood urea nitrogen (BUN) and creatinine (Cr) levels were assessed by GC Labs (Yongin, Republic of Korea) using a Cobas 8000 modular analyzer system (Roche, Germany).
Kidney tissues from each experimental group were fixed with 4% paraformaldehyde (pH 7.4) and embedded in paraffin. 2-µm tissue sections were prepared and stained with Masson’s trichrome using standard protocols to determine histological changes and collagen deposition. Collagen deposition was quantified as a percentage of the total area using iSolution DT image software (IMT iSolution, Vancouver, Canada) in more than nine randomly selected fields in the cortex and medulla sections. For immunohistochemical staining, 2-μm-thick kidney sections were deparaffinized and rehydrated, and endogenous peroxidase was inactivated with 3% hydrogen peroxide. The samples were incubated with anti-CD31 (1:200; ab182981, Abcam) overnight at 4 °C and then detected using the EnVision-HRP kit (Dako, Carpinteria, CA, USA). To quantify CD31 expression, the marked cells were counted in nine non-overlapping fields imaged at 400 × magnification and divided by the area of the field of view for each slide.
Kidney tissues from each experimental group were fixed with 4% paraformaldehyde (pH 7.4) and embedded in paraffin. 2-µm tissue sections were prepared and stained with Masson’s trichrome using standard protocols to determine histological changes and collagen deposition. Collagen deposition was quantified as a percentage of the total area using iSolution DT image software (IMT iSolution, Vancouver, Canada) in more than nine randomly selected fields in the cortex and medulla sections. For immunohistochemical staining, 2-μm-thick kidney sections were deparaffinized and rehydrated, and endogenous peroxidase was inactivated with 3% hydrogen peroxide. The samples were incubated with anti-CD31 (1:200; ab182981, Abcam) overnight at 4 °C and then detected using the EnVision-HRP kit (Dako, Carpinteria, CA, USA). To quantify CD31 expression, the marked cells were counted in nine non-overlapping fields imaged at 400 × magnification and divided by the area of the field of view for each slide.
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