FGG and IGKC were analysed by ELISA using commercially available kits (Kappa Human ELISA kit from Abcam ab157709, lot: GR3174712-5; Fibrinogen Human ELISA kit from Abcam, ab108841, lot: GR3177851-6) in a subset of the individuals to confirm the pattern of variation observed in the proteome analysis. The analyses were performed as described in the protocol.
Human fibrinogen elisa kit
Manufactured by Abcam
Sourced in Japan, United Kingdom
This Human fibrinogen ELISA kit is designed to quantitatively measure human fibrinogen levels in biological samples. It employs the quantitative sandwich enzyme immunoassay technique.
Automatically generated - may contain errors
Lab products found in correlation
Ab108842, by Abcam (1 mentions)
Human tnf α quantikine elisa, by R&D Systems (1 mentions)
Serum triglyceride determination kit, by Merck Group (1 mentions)
Ranbut, by Randox (1 mentions)
Vacuette, by Greiner (1 mentions)
Human pmn elastase elisa kit, by Abcam (1 mentions)
Simplestep elisa kit, by Abcam (1 mentions)
Quantikine elisa kit, by R&D Systems (1 mentions)
Human elisa kit, by Merck Group (1 mentions)
Glomax discover microplate reader, by Promega (1 mentions)
5 protocols using human fibrinogen elisa kit
1
Validating Protein Expression via ELISA
2
Postprandial Metabolic and Inflammatory Markers
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At each time point, 10 mL of venous whole blood was taken and dispensed into serum separation and lithiumheparin (Vacuette; Greiner Bio-One GmbH, Kremsmünster, Austria) tubes before being centrifuged for 15 min at 2000g at 4°C and stored at -80°C for retrospective analysis of triglycerides (TGs) (Serum Triglyceride Determination Kit; Sigma-Aldrich, St. Louis, MO, USA). Apolipoprotein B48 (APO B48 ; Antibodies-online, USA), non-esterified-fatty acids (NEFA) (RANBUT; Randox Laboratories, London, UK), plasma glucagon (Glucagon EIA; Sigma-Aldrich) and tumour necrosis factor alpha (TNF-α) (Human TNF-α Quantikine ELISA; R&D Systems, Roche Diagnostics, West Sussex, UK) were measured hourly. Plasma fibrinogen (ab108842, Fibrinogen Human ELISA Kit; Abcam, Japan), HTF (Human Tissue Factor activity ab108906; Abcam, UK) and PAI-1 (Human PAI-1/serpin ELISA Kit DSE100; R&D systems, UK) were measured at rest, 3 h and 6 h postmeal. The intra-assay coefficient of variation was <10% for all assays. Due to increased assay cross-reactivity with insulin detemir, only participants treated with insulin glargine were included in serum insulin analysis (n = 8).
3
Fibrinogen Adsorption on PA66 Discs
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PA66 discs were utilized as substrates and results of bare discs were listed as control. Human fibrinogen ELISA kit was purchased from Abcam Co. Ltd. Fresh human whole blood was acquired from healthy human volunteers, 3.2% trisodium citrate was added with the volume ratio of 1:9 followed by centrifugal for 15 min at 3000 rpm to prepare platelet poor plasma (PPP). 100 μL PPP was added onto the samples, negative and positive control groups, and incubated for 2 h at 37 °C. Human fibrinogen ELISA kit was utilized to measure adsorbed Fg.
4
Biomarker Measurement using ELISAs
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The levels of five biomarkers were measured using ELISAs. The plasma stored at −79°C was defrosted in a constant temperature water bath to 37°C. CRP (Human C-Reactive Protein/CRP Immunoassay kit; cat. no. DCRP00), CC16 (Human Uteroglobin Quantikine ELISA kit; cat. no. DUGB00) and TGF-β (Human TGF-beta Quantikine ELISA kit; cat. no. DB100B) levels were assessed using Quantikine ELISA kits (all R&D Systems, Inc.), while fibrinogen(Human Fibrinogen ELISA kit; ab208036) and neutrophil elastase (Human PMN Elastase ELISA kit; ab119553) were analyzed using Simple Step ELISA kits (Abcam) according to the to the instructions of the respective manufacturer's instructions.
5
Quantifying Fibrinogen and Growth Factors in PRP
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Fibrinogen (FIB) concentration (μg/mL) was quantified with Human Fibrinogen ELISA Kit (ab108842, Abcam, Cambridge, UK), following the manufacturer’s instructions. Each measure was performed on three different PRP replicates of the same unit and assessed in triplicate. GFs (Table 3 ) concentration was assessed by the following Human ELISA kits (Sigma-Aldrich s.r.l., Milan): PDGF-AA (RAB0394), PDGF-BB (RAB0397), PDGF-AB (RAB0396), EGF (RAB0149). Each measure was performed on four different PRP dilutions, except for the PDGF-AA (three dilutions), and assessed in duplicate. ELISA 96-well plates were read with Glomax® Discover Microplate Reader (Promega, Madison, WI, USA).
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