Serum levels of ANGPTL8 were determined by using the ELISA kit (enzyme-linked immunosorbent assay kit) for ANGPTL8 (Cloud-Clone Corp., TX, USA) according to the manufacturer's instructions. Detection limit of the kit was 0.056 ng/mL. Samples were 30-fold diluted. The absorbance values were read at 450 nm on a Multiskan RC ELISA reader (Thermo Fisher Scientific, Waltham, MA, USA).
Multiskan rc elisa reader
Manufactured by Thermo Fisher Scientific
Sourced in United States
The Multiskan RC ELISA reader is a microplate reader designed to measure absorbance in microplates. It is capable of performing spectrophotometric analysis and can be used for various ELISA-based applications.
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18 protocols using multiskan rc elisa reader
1
ANGPTL8 Serum Levels Quantification
2
Quantification of Anti-oxLDL Antibodies
3
Quantification of Cervical IL-6 Levels
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The cervical fluid concentration of IL-6 was assessed by the Human IL-6 Quantikine enzyme-linked immunosorbent assay (R&D Systems Inc., Minneapolis, MN, USA). The minimal detectable concentration was 0.70 pg/mL, and the interassay and intraassay coefficients were less than 10%. Absorbance values were measured at 450 nm using a Multiskan RC ELISA reader (Thermo Fisher Scientific, Waltham, MA, USA). AF IL-6 concentrations were assessed with a lateral flow immunoassay Milenia® QuickLine IL-6 using Milenia® POCScan Reader (Biotec GmbH, Giessen, Germany). The measurement range was 50–10000 pg/mL. Intra-lot- and inter-lot-variation was 12.1% and 15.5%, respectively.
4
Cytokine Profiling by ELISA
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The promising results obtained from the screening examination were subsequently verified by a more sensitive enzyme-linked immunosorbent assay technique (ELISA) using ELISA kits for human IL-1α, IL-1ra, IL-6, and VEGF (R&D systems, Minneapolis, MN, USA) according to the manufacturer’s instructions. The minimal detectable doses of selected kits were 0.1 pg/mL for IL-1α, 2.2 pg/mL for IL-1ra, 0.7 pg/mL for IL-6, and 5.0 pg/mL for VEGF. Absorbance was read at 450 nm using a Multiskan RC ELISA reader (Thermo Fisher Scientific, Waltham, MA, USA). All values were expressed in pg per milliliter of the serum.
5
Biomarker Measurement with ELISA
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Selected biomarkers were measured with commercial ELISA kits according to the manufacturer’s instructions and the absorbance values were read at 450 nm on a Multiskan RC ELISA reader (Thermo Fisher Scientific, Waltham, MA, USA).
6
CD200R1 Levels ELISA Quantification
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CD200R1 levels were determined using an ELISA kit: ELISA Assay Kit for CD200 Receptor 1 (Cloud Clone Corp., Katy, TX, USA) according to the manufacturer’s instructions. Samples were diluted 100-fold. The detection range was 7.8–1000 pg/mL. The absorbance values were read at 450 nm on a Multiskan RC ELISA reader (Thermo Fisher Scientific, Waltham, MA, USA).
7
Quantifying Anti-PCP Antibodies by ELISA
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The serum levels of anti-PCP were determined by ELISA using specific VaccZyme™ ELISA kits for human anti-PCP IgG, IgG2, and IgA (Binding Site, Birmingham, UK), according to the manufacturer's instructions. The range of detection was 3.3–270 mg/L for the IgG kit, 1.1–90 mg/L for the IgG2 kit, and 0–270 U/mL for the IgA kit. Serum samples were diluted 100x, and the absorbance was read at 450 nm using a Multiskan RC ELISA reader (Thermo Fisher Scientific).
8
Quantification of Anti-oxLDL Antibodies
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These parameters were obtained by fractionation of LDL and preparation of oxLDL as antigen in the antibody detection assay, according to what reported by Ketelhuth et al. [29 (link)]. For this purpose a commercially available ELISA kits (ChusaBiotech Co. Ltd., China), was employed. Briefly, microtiter plates were coated with 10 g/ml in PBS of either native LDL or MDA-LDL to measure anti-oxidized LDL antibodies. Serum samples were diluted 1:100 in 1% BSA/PBS and incubated for 2 h at 37°C and overnight at 4 C. Afterwards, the plates were washed four times with alkaline phosphatase-conjugated anti-human IgG (Sigma Chemical, St. Louis, MO) and left for 3 h at room temperature. The reaction was halted after 60 min with 1% BSA/PBS for 2 h at room temperature. The absorbance was at 450 nm using a Multiskan RC ELISA reader (Thermo Fisher Scientific, USA). The binding of antibodies to oxidized LDL was determined by subtracting native LDL binding value from the one of MDA-LDL while the results were expressed as an Optical Density (OD).
9
Anti-αGal Antibody Quantification by ELISA
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The serum concentration of anti-αGal was determined by a sandwich enzyme-linked immunosorbent assay (ELISA), using an ELISA kit for human anti-alpha galactosyl IgG, IgA, and IgM (BioVendor, Laboratorní Medicína a.s., Brno, Czech Republic) according to the manufacturer's instructions. The range of detection was 3.13–100 U/mL for every kit. Serum samples were diluted 100x, and absorbance was read at 450 nm using a Multiskan RC ELISA reader (Thermo Fisher Scientific, Waltham, MA, USA).
10
Elafin Serum Level Quantification
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Serum levels of Elafin were determined using the ELISA kit Elafin/Skalp Human ELISA Kit (Abcam, UK), according to the manufacturer’s instructions. The sensitivity of the kit was 5 ng/mL. Samples were 150-fold diluted. The absorbance values were read at 450 nm on a Multiskan RC ELISA reader (Thermo Fisher Scientific, Waltham, MA, USA).
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