Ultra sensitive enzyme linked immunosorbent assay

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The Ultra-sensitive enzyme-linked immunosorbent assay (ELISA) is a laboratory technique used for the detection and quantification of specific proteins or other biomolecules in a sample. It employs antibodies and color change to identify and measure the target analyte. The ultra-sensitive ELISA offers enhanced detection capabilities compared to standard ELISA methods.

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Lab products found in correlation

Highly sensitive immunoturbidimetric assay, by Denka Seiken (2 mentions) Enzyme linked immunosorbent assay, by R&D Systems (2 mentions) Bnii nephelometer, by Siemens (2 mentions) Radioimmunoassay, by Linco Research (1 mentions)

8 protocols using ultra sensitive enzyme linked immunosorbent assay

Plasma 25(OH)D and Colorectal Cancer: Inflammatory Markers

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To account for the potential confounding effect by systemic inflammation on the plasma 25(OH)D-CRC association, we also measured three inflammatory markers in our study samples: C-reactive protein (CRP), interleukin 6 (IL6), and tumour necrosis factor receptor superfamily member 1B (TNFRSF1B, also known as soluble tumour necrosis factor receptor 2, sTNFR-2). We used a highly sensitive immunoturbidimetric assay (Denka Seiken Co, Tokyo, Japan) to measure CRP levels, an ultra-sensitive enzyme-linked immunosorbent assay (R&D Systems, Minneapolis, MN) to measure IL6, and an enzyme-linked immunosorbent assay (R&D Systems, Minneapolis, MN) to measure TNFRSF1B levels. More details regarding the measurements can be found in previous publications.[20 (link), 21 (link)]

Song M., Nishihara R., Wang M., Chan A.T., Qian Z.R., Inamura K., Zhang X., Ng K., Kim S.A., Mima K., Sukawa Y., Nosho K., Fuchs C.S., Giovannucci E.L., Wu K, & Ogino S. (2015). Plasma 25-hydroxyvitamin D and colorectal cancer risk according to tumour immunity status. Gut, 65(2), 296-304.

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Plasma Galectin-3 Levels and Stroke Risk

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Fasting blood samples were drawn at the baseline home visit for each subject using previously published standardized methods [10 (link)]. Plasma galectin-3 was measured in the case–cohort sample using an ultra-sensitive enzyme-linked immunosorbent assay (R&D Systems, Minneapolis, MN, USA). The laboratory analytical coefficient of variation range was 5.6%–7.4%. The lower detection limit was 1.8 ng/ml. Of the 1453 participants, two (0.1%) had values below the lower limit. A value halfway between 0 and the lower limit was assigned to these, i.e. 0.9 ng/ml. The upper detection limit was 35 ng/ml. There were two (0.1%) participants with values above the upper limit. For these values 10% higher than the upper limit, i.e. 38.5 ng/ml, was assigned.
There were 111 participants with missing galectin-3 (8% of cases and 7% of the CRS), mostly due to missing samples. This left 526 stroke cases (including 20 who were members of the CRS) and 947 in the CRS.

Arora P., Agarwal Z., Venkatraman A., Callas P., Kissela B.M., Jenny N.S., Judd S.E., Zakai N.A, & Cushman M. (2017). Galectin-3 and risk of ischaemic stroke: Reasons for Geographic and Racial Differences in Stroke cohort. European journal of neurology, 24(12), 1464-1470.

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Biomarker Measurements in Clinical Research

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CRP (mg/L) was measured by nephelometry (BNII nephelometer, Dade
Behring, Deerfield, IL). The intra-assay and inter-assay analytical
coefficients of variation for CRP ranged from
2.3%–4.4% and from
2.1%–5.7%, respectively. IL-6 (pg/mL) was measured
by ultrasensitive enzyme-linked immunosorbent assay (R&D Systems,
Minneapolis, MN) with an analytical coefficient of variation of
6.3%. The distributions of both IL-6 and CRP were highly skewed, so
they were log-transformed for the analyses.

Kershaw K., Lewis T.T., Diez Roux A.V., Jenny N.S., Liu K., Penedo F.J, & Carnethon M. (2016). Self-reported experiences of discrimination and inflammation among men and women: The Multi-Ethnic Study of Atherosclerosis. Health psychology : official journal of the Division of Health Psychology, American Psychological Association, 35(4), 343-350.

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Inflammatory Biomarkers Assessment Methodology

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Two markers of inflammation were used in the analysis: C-reactive protein (CRP) and interleukin-6 (IL-6). CRP was initially measured from plasma using a particle enhanced immunonephelometric assay (BNII nephelometer; Dade Behring Inc., Deerfield, IL). The intra- and inter-assay coefficients of variance (CVs) ranged from 2.3 % to 4.3 % and 1.1–4.4 %, respectively. The final batch of samples (collected after February 1, 2015; N = 393) as well as those below the limit of detection using the immunonephelometric assay (N = 27) were assayed from serum using the Meso Scale Diagnostics (MSD) immunoelectrochemiluminescent platform (intra-assay CV: 2.2–4.1 %; inter-assay CV: 4.7–5.2 %). The data from the two platforms was integrated using the data adjustment formulas described in the study documentation (see Weinstein et al., 2018 (link) for details). IL-6 was measured from serum using an ultrasensitive enzyme-linked immunosorbent assay (R&D Systems, Minneapolis, MN). The intra-assay CV was 4.7 % and the inter-assay CV ranged from 5 % to 15 %. CRP and IL-6 values were log-transformed to achieve a normal distribution. One participant had missing data for IL-6 and 4 participants had missing data for CRP due to an insufficient amount of blood collected and were thus not included in the analyses.

Lee D.S, & Way B.M. (2021). Social media use and systemic inflammation: The moderating role of self-esteem. Brain, Behavior, & Immunity - Health, 16, 100300.

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Measurement of Plasma Biomarkers in Epidemiological Studies

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We used the radioimmunoassay method to measure plasma 25(OH)D, a highly sensitive immunoturbidimetric assay (Denka Seiken Co, Tokyo, Japan) to measure CRP, an ultra-sensitive enzyme-linked immunosorbent assay (R&D Systems, Minneapolis, MN) to measure IL-6, and an enzyme-linked immunosorbent assay (R&D Systems, Minneapolis, MN) to measure soluble tumor necrosis factor receptor 2 (sTNFR-2), as previously described (5 (link), 14 (link), 15 ). Samples from cases and their matched controls were handled together and analyzed in the same batch. Quality control samples were randomly interspersed among the case-control samples. Personnel blinded to quality control and case-control status conducted all assays. For 25(OH)D, the mean intra-assay coefficient of variation from blinded quality control samples was <15% for all batches. For inflammatory markers, the intra-assay coefficients of variations in the NHS and HPFS were as follows: CRP, 2.2% and 7.8%; IL-6, 10.6% and 12.1%; sTNFR-2, 6.7% and 10.1% (14 (link), 15 ). We excluded plasma samples that failed any of the four laboratory assays from 19 cases and 28 controls in the NHS, and 13 cases and 19 controls in the HPFS.

Song M., Wu K., Chan A.T., Fuchs C.S, & Giovannucci E.L. (2014). Plasma 25-hydroxyvitamin D and risk of colorectal cancer after adjusting for inflammatory markers. Cancer epidemiology, biomarkers & prevention : a publication of the American Association for Cancer Research, cosponsored by the American Society of Preventive Oncology, 23(10), 2175-2180.

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Inflammatory Biomarker Measurement Protocol

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Inflammatory biomarkers were previously measured in a WHI ancillary study conducted in a sample of women in the WHI-OS at baseline from 1993 to 1998 (n=3,245) (22 (link)). The construct validation of the DII was conducted in this sample of women who had data on plasma IL-6, hs-CRP and TNFα-R2. We also derived an overall inflammatory biomarker score by standardizing each of the three inflammatory biomarkers as described under statistical analysis.
Methods for the collection and processing of blood samples have been described (22 (link)). Briefly, IL-6 and TNFα-R2 concentrations were measured by an ultrasensitive enzyme-linked immunosorbent assay (R&D Systems, Minneapolis, Minnesota), and hs-CRP concentrations were measured on a chemistry analyzer (Hitachi 911; Roche Diagnostics, Indianapolis, Indiana) using an immunoturbidimetric assay with reagents and calibrators (Denka Seiken Co. Ltd., Niigata, Japan). The coefficients of variation were: 7.6% for IL-6,1.6% for hs-CRP, and 3.5% for TNFα-R2 (22 (link)).

Tabung F.K., Steck S.E., Zhang J., Ma Y., Liese A.D., Agalliu I., Hingle M., Hou L., Hurley T.G., Jiao L., Martin L.W., Millen A.E., Park H.L., Rosal M.C., Shikany J.M., Shivappa N., Ockene J.K, & Hebert J.R. (2015). Construct Validation of the Dietary Inflammatory Index among Postmenopausal Women. Annals of epidemiology, 25(6), 398-405.

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Cytokine and Inflammation Biomarker Assays

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Blood samples were collected in the morning when participants underwent venipuncture at the baseline visit after an overnight fast, and serum samples were frozen at -70°C. Both IL-6 and TNF- α were measured in duplicate using an ultrasensitive enzyme-linked immunosorbent assay (R&D Systems, Minneapolis, MN, USA). The limit of detection was 0.10 pg/ml for IL-6 and 0.18 pg/ml for TNF- α. The level of PAI-1 was measured in citrated plasma samples using a 2-site enzyme-linked immunosorbent assay. Serum levels of CRP were also measured in duplicate by enzyme-linked immunosorbent assay based on purified protein and polyclonal anti-CRP antibodies (Calbiochem, San Diego, CA, USA) with a coefficient of variation of 8.0%. Serum concentrations of leptin were measured in duplicate by means of radioimmunoassay (Linco Research Inc, St Charles, MO, USA). The assay range is 0.05 to 100 ng/ml leptin in serum. Intra-assay CVs ranged from 3.7% to 7.5%, and inter-assay CVs ranged from 3.2% to 8.9%.

Mishra S., Harris T.B., Hsueh W.C., Hue T., Leak T.S., Li R., Mehta M., Vaisse C, & Sahyoun N.R. (2015). The Association of Serum Leptin with Mortality in Older Adults. PLoS ONE, 10(10), e0140763.

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Plasma Biomarkers Assessment Methods

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Plasma fibrinogen was assessed by the Clauss method in 1990 and Behring Nephelometer II (BN II) method (Dade Behring, Deerfield, IL) in 1992–1993 and 2005–2006, calibrated with standard normal plasma [27 (link)]. Intra- and inter-assay CVs were 2.3% and 4.4%, respectively, in 1992–1993; and 3.1% and 4.2%, respectively, in 2005–2006. Plasma high-sensitivity C-reactive protein (hs-CRP) was measured from blood samples collected in 1992–1993, 2000–2001, 2005–2006 and 2010–2011 utilizing a particle enhanced immunonephelometric assay [28 (link)]. Intra-assay CVs ranged from 2.3% to 4.4%, and inter-assay CVs from 2.1% to 5.7% across different examinations. Plasma interleukin-6 (IL-6) was analyzed in 2005–2006 only by ultra-sensitive enzyme-linked immunosorbent assay (R&D Systems, Minneapolis, MN), with a routine inter-assay CV of 6.3% [29 (link)].

Tang W., Xun P., Chen C., Lu L., Sood A., Shikany J.M, & Kahe K. (2020). Association between toenail zinc concentrations and incidence of asthma among American young adults: the CARDIA study. Journal of trace elements in medicine and biology : organ of the Society for Minerals and Trace Elements (GMS), 64, 126683.

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