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Luminex flow cytometry

Manufactured by Bio-Rad
Sourced in United States

The Luminex flow cytometry product is a versatile tool for multi-analyte analysis. It utilizes color-coded magnetic beads to enable simultaneous detection and quantification of multiple target analytes in a single sample. The core function of the Luminex system is to facilitate high-throughput, multiplexed assays for applications such as protein and nucleic acid detection.

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7 protocols using luminex flow cytometry

1

Abdominal Aortic Calcium and Adipokines

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A random subset of all MESA participants (n = 1,970) underwent abdominal computed tomography (CT) scans at either exam 2 or exam 3 (randomly assigned) to measure abdominal aortic calcium (40 (link)). In a subsequent ancillary study related to body composition, adipokine levels were measured among these same participants from stored frozen serum samples obtained at the respective exams (2 or 3) of their CT scan (30 (link)). The adipokines (adiponectin, leptin, and resistin) were analyzed using a Bio-Rad Luminex flow cytometry (Millepore, Billerica, MA) at the Laboratory for Clinical Biochemistry Research (University of Vermont, Burlington, VT), as previously reported (31 (link), 32 (link)). The coefficients of variation for these assays ranged from 6 to 13%.
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2

Metabolic Biomarkers in Cardiometabolic Health

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Total and HDL cholesterol, triglycerides, and glucose levels were measured from blood samples obtained after a 12-hour fast [21 (link)]. Diabetes was defined as fasting glucose ≥ 126 mg/dL or use of hypoglycemic medication. Fasting glucose between 100–125 mg/dL was considered impaired fasting glucose. Stored fasting blood samples were analyzed to provide levels of adiponectin, leptin, TNF-α and resistin. These adipokines were measured using Bio-Rad Luminex flow cytometry (Millepore, Billerica, Massachusetts) at the Laboratory for Clinical Biochemistry Research (University of Vermont, Burlington, Vermont). Average analytical coefficients of variation across several control samples for these analytes ranged from 6.0–13.0%.
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3

Metabolic Biomarkers in Clinical Research

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At each visit, 12hr-fasting venous blood samples were obtained and processed using standard methods.(11 (link)) Total and high-density lipoprotein cholesterol, triglycerides, insulin and glucose levels were measured. Samples from clinic visits 2 and 3 were assayed for IL-6, resistin, CRP, and TNF-α as well as the adipokines leptin and adiponectin. CRP was measured by immunonephelometry using the BNII instrument (N High Sensitivity CRP, N Antiserum to Human Fibrinogen, Dade Behring Inc., Deerfield, Ill), while IL-6 was measured by ultrasensitive ELISA (Quantikine HS Human IL-6 Immunoassay; R&D Systems, Minneapolis, Minn). Resistin, TNF-α, leptin, and adiponectin concentrations were measured using Bio-Rad Luminex flow cytometry (Millepore, Billerica, MA, USA). These analyses were performed at the Laboratory for Clinical Biochemistry Research (University of Vermont, Burlington, VT). Participants who used cholesterol reducing medication or with a total cholesterol/high-density lipoprotein cholesterol ratio > 5.0 were classified as dyslipidemic, whilst those who used hypoglycemic medication or with a fasting glucose ≥ 126 mg dl−1 were classified as diabetic.
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4

Measuring Obesity-Related Adipokines and Insulin

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As previously reported,(16 (link),18 (link)) the obesity-related adipokines (adiponectin, leptin, and resistin), were measured from stored (fasting) blood from the CT visit (visit 2 or visit 3) using a Bio-Rad Luminex flow cytometry (Millepore, Billerica, MA) at the Laboratory for Clinical Biochemistry Research (University of Vermont, Burlington, VT). The coefficients of variation (CV) ranged from 6 to 13%. Insulin was measured by radioimmunoassay using the Linco Human Insulin Specific assay (Linco Research, Inc., St. Charles, MO), with CV of 4.9%. The adipokine and insulin biomarkers had a skewed distribution and were log-transformed for all analyses.
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5

Adipokines Measurement from Frozen Serum

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Fasting serum samples were obtained at either visit 2 (2002-2004) or visit 3 (2004-2005) and frozen at −70°C (14 (link)). In 2009, adipokines (adiponectin, leptin and resistin) were measured from these stored frozen serum samples using a Bio-Rad Luminex flow cytometry at the Laboratory for Clinical Biochemistry Research (14 (link)). The adipokine assays were validated against several gold standard ELISAs and the average analytical coefficients of variation across several control samples ranged from 6.0-13.0% (14 (link)).
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6

Biomarker Measurements in MESA Cohort

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At each clinic visit, venous blood was collected after a 12-hour fast and shipped to the MESA central laboratory for analysis of total and HDL cholesterol, triglycerides, glucose, and creatinine concentration. Stored fasting blood samples from visits 2 or 3 were analyzed for insulin, CRP, fibrinogen, renin, aldosterone, adiponectin, leptin, TNF-α, IL-6 and resistin. Adipokines were measured using Bio-Rad Luminex flow cytometry (Millepore, Billerica, MA) at the laboratory for Clinical Biochemistry Research (University of Vermont, Burlington, VT). Average analytic coefficients of variation across several control samples ranged from 6.0% to 13.0%. Dyslipidemia was defined as a total cholesterol/HDL-cholesterol ratio >5.0 or if the participant was taking medication to reduce cholesterol. Diabetes was defined as fasting glucose ≥ 126 mg·dL−1 or use of hypoglycemic medication. Estimated glomerular filtration rate (eGFR) was calculated using the chronic kidney disease-Epi equation (19 (link)).
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7

Biomarker Measurements in Fasting Blood

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At visits 2 and 3, venous blood was collected after a 12-hour fast, then shipped to the MESA central laboratory for the measurement of total cholesterol, high-density lipoprotein (HDL) cholesterol and glucose levels. Dyslipidemia was defined as a total cholesterol to HDL cholesterol ratio >5.0 or the use of any lipid-lowering medication. Fasting blood was also used for the measurement of insulin levels and the inflammation markers C-reactive protein (CRP), fibrinogen and IL-6 as described previously [15 (link), 17 (link)]. Circulating levels of adiponectin, leptin, TNF-α, and resistin were measured in stored fasting blood samples from visits 2 and 3 using Bio-Rad Luminex flow cytometry (Millepore, Billerica, MA) [15 (link)].
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