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ELISA

Manufactured by Abnova
Sourced in Taiwan, Province of China, United States

ELISA (Enzyme-Linked Immunosorbent Assay) is a biochemical technique used to detect and quantify specific substances, typically proteins, antibodies, or other biomolecules, in a liquid sample. The core function of ELISA is to measure the presence and concentration of a target analyte through the use of antibodies and color-change or fluorescent reactions.

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54 protocols using ELISA

1

Norepinephrine Levels in Hippocampus and Plasma After Conditioning

Animals in which the dorsal hippocampus was injected with propranolol (5 μg per side) were sacrificed either under basal conditions or 30, 60, and 120 min after conditioning with 1.4‐mA shocks. Trunk blood and the dorsal hippocampus were quickly collected from animals subjected to each of the conditions. Rat hippocampal samples were dissected on dry ice and homogenized on ice to generate lysates (1 mg wet weight tissue into 40 μl of 0.01 N HCl, 1 mmol/L EDTA, and 4 mmol/L sodium metabisulfite). After centrifugation of blood in EDTA‐coated tubes at 2000 rpm for 10 min, the supernatant was stored at −80°C until the assay was performed. Norepinephrine was extracted from brain tissues according to the manufacturer's instructions, and the dried extracts were stored at −80°C until the assay was performed. Plasma and hippocampal levels of norepinephrine or epinephrine were determined by specific enzyme‐linked immunosorbent assays (ELISAs; Abnova, Taipei, Taiwan), according to the manufacturer's instructions.
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Lipopolysaccharide binding protein (LBP) and plasminogen activator inhibitor-1 (PAI-1) protein levels were measured in peripheral plasma of humans and plasma from portal vein of mice using commercially available ELISAs (Abnova, Taiwan and LOXO, Germany), respectively, as detailed by the manufacturer. Arginase activity in proximal small intestine was measured, as previously described [21 (link)].
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LBP was quantified from clarified and 800-fold diluted fasting heparin plasma with ELISAs (Abnova, Taipei City, Taiwan; cat number KA0448) as described elsewhere [28 (link)].
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Saliva was collected at the first 2 h, 7th and 14th days of life. The saliva samples on the 7th and 14th days after birth were collected 2 h after tube-feeding in the morning. The samples were centrifuged at 5000 rpm for 20 min and the supernatant were stored in freeze (− 30 °C) until analysis. Before measurement, collection tubes were removed from the freezer and thawed at room temperature for 10–15 min. The concentration of sIgA in the samples were then measured by quantitative enzyme-linked immunosorbent assay (ELISA) (Abnova, Taiwan, China) following the protocol provided by the manufacturer. After the samples were stained, the absorbance (OD value) was measured at 450 nm, then the concentration of sIgA was calculated. The instrument involved in the experiment was the enzyme-label measuring instrument (Muitiskan FC, Shanghai, China).
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HepG2 cells stably expressing human NTCP-myc in a 96-well plate were treated with preS1 peptide, anti-ovalbumin (anti-OVA) MAb, or N6HB426-20 MAb for 1 h and then washed with PBS. Cells were infected with HBV at a concentration of 500 genome equivalents per cell in the presence of 4% PEG 8000 and 2% dimethyl sulfoxide (DMSO) along with the preS1 peptide or each antibody. At 24 h postinfection, cells were washed with PBS and cultured in fresh medium containing 2% DMSO. At 12 days postinfection, the level of HBeAg was determined by enzyme-linked immunosorbent assay (ELISA; Abnova, Taipei, Taiwan).
Alternatively, PXB cells were treated with preS1 peptide, anti-OVA MAb, or N6HB426-20 MAb for 1 h and washed with PBS. Cells were infected with HBV (genotype C or genotype D derived from HBV-infected patients) at a concentration of 10 genome equivalents per cell in the presence of 4% PEG 8000 and 2% DMSO, along with preS1 peptide or each antibody. At 24 h postinfection, cells were washed with PBS. HBV-infected cells were cultured in fresh medium containing 2% DMSO. At day 7 postinfection, the level of HBsAg was determined by a chemiluminescent enzyme immunoassay (Lumipulse f, Fujirebio, Japan).
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Detection and quantification of HbeAg levels was performed by ELISA according to the manufacturer’s instructions (Abnova, Taipei, Taiwan). Briefly, a 100 ul sample of 1:10 diluted supernatant was used in lieu of undiluted supernatant. Absorbance was read at 450λ on the BertholdTech TriStar (Bad Wildbad, Germany).
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Plasma was collected at 96 hours post MCAo. Lipopolysaccharide binding protein (LBP) levels were determined by ELISA (ABNOVA) following manufacturer’s instructions. Plasma cytokines [tumor necrosis factor (TNF) α, interleukin-12 (IL-12), interleukin-6 (IL-6), interleukin-1 (IL-1) alpha, interleukin-13 (IL-13) and chemokine ligand-1 (CXCL1)] were determined using multiplex (BIORAD) and were run in duplicates. Inter-and intra-assay coefficients of variation were less than 10%.
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Red blood cell membranes were collected from healthy human and coated with nanoparticles (RNP) following previous methods [28 (link)]. The RNP and CD90@NP were labeled with DiD (5 μM, Fanbo Biochemicals Co. Ltd, Beijing, China) at the concentration of 0.2% (w/w) for 10 min. Triamcinolone acetonide (18026, Cayman Chemical Company, MI, USA)-loaded NP (T-NP), RNP (T-RNP), and CD90@NP (T-CD90@NP) were prepared by adding 20% (w/w) TA to the respective solutions during the PLGA core preparation.
The structure micro humid of the CD90@NP was observed using the HT7800 transmission electron microscope (Hitachi Ltd, Tokyo, Japan). The ζ potential (mA) and particle size (nm) of the CD90@MV, NP, RNP, and CD90@NP were detected by a Malvern Zetasizer Nano ZS90 nanoparticle size system (Malvern, UK). T-CD90@NP was collected via centrifugation at 12,000×g for 45 min and dissolved in dimethyl sulfoxide to determine the DLC and EE of TA. The TA content in T-CD90@NP was tested by ELISA (Abnova, Taipei, Taiwan). The following formula was used to calculate the DLC and EE: DCL%=TAencapsulatedinT - CD90@NPweightofT-CD90@NP×100%, EE%=TAencapsulatedinT-CD90@NPtotalTA×100%.
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9

Cytokine, Intestinal Barrier, and Bacterial Translocation Analysis

To study the concentrations of IL-10 in the serum, the Milliplex MAP Kit (MERCK, Darmstadt, Germany) was employed using the protocol recommended by MERCK. The plate was read in a Luminex MAGPIX with xPONENT software. The concentration of the cytokine was calculated from the standard curve using mean fluorescence intensity (MFI) analysis with Analyst software (MERCK).
To study intestinal barrier damage, the concentration of I-FABP and zonulin in the serum were measured by enzyme-linked immunosorbent assay (ELISA). I-FABP was purchased from Hycult Biotech (Hycult Biotech, PA, USA), and zonulin was purchased from R&D Systems (R&D Systems, MN, USA). #e tests were carried out according to the protocols provided in the kits. #e plate was read in an iMark Microplate Reader at 450 nm with Microplate Manager So(ware (Termo Fisher Scientific, MA, USA).
To study bacterial translocation, the concentration of LBP in the serum was measured by ELISA (Abnova, Taipei, Taiwan). We performed 1:800 dilutions of the samples from the MDD patients and HCs. The test was carried out according to the protocol provided in the kit. The plate was read in an iMark Microplate Reader at 450 nm with Microplate Manager Software (Thermo Fisher Scientific).
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Plasma PSA was measured by ELISA (Abnova) as per manufacturer’s instructions using FLUOstar Omega spectrophotometer.
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