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AssayMax ELISA kits

Manufactured by Assaypro
Sourced in United States

AssayMax ELISA kits are a quantitative enzyme-linked immunosorbent assay (ELISA) designed for the detection and quantification of specific analytes in biological samples. The kits provide a reliable and accurate method for measuring target molecules in a wide range of sample types.

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9 protocols using AssayMax ELISA kits

To assess the APR protein secretion, culture media from primary human hepatocyte cultures were collected over the final 24 h of treatment, flash-frozen in liquid nitrogen, and stored at -80°C prior to analysis. Secreted proteins were detected with AssayMax™ ELISA Kits (AssayPro).
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Quantification of four interesting targets; hemopexin (HPX), apolipoprotein E and A-I (Apo E and A-I), and plasma retinol binding protein (RBP4) was performed by using ELISA. Concentrations of the candidate proteins in serum samples of different grades of meningiomas; grade I (n = 14), grade II (n = 5) and grade III (n = 1) and healthy controls (n = 45) were measured using AssayMax ELISA kits following the manufacturer's instructions (AssayPro, USA). Optical densities were measured at 450 nm and 570 nm using a SpectraMax M2e (Molecular Devices, USA). Statistical significance of differential expression of the serum proteins was analyzed by Mann Whitney U-test.
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One day prior to the aforementioned cognitive assessments, all participants were instructed to follow the protocol modified from the work of McLean, et al.36 (link) to collect their saliva in sample tubes at home at four time points: awakening, 30 min after awakening, 3 pm, and bedtime. For saliva sampling at awakening, participants were asked not to brush their teeth, eat, or drink before sampling. All collected saliva samples were brought to the hospital the next day and preserved in our laboratory refrigerator at − 20 °C until further analysis using commercial enzyme-linked immunosorbent assay kits, AssayMax ELISA Kits (Assaypro, St. Charles, MO, USA). The cortisol awakening response (CAR) was calculated as the difference in cortisol levels between awakening and 30 min after awakening37 (link).
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Five selected targets namely, Serum amyloid A (SAA; P0DJI8), Hemopexin (HPX; P02790), Apolipoprotein E (Apo E; P02649), Haptoglobin (HP; P00738), and Apolipoprotein A1 (Apo A1; P02647) were quantified using AssayMax ELISA kits (AssayPro, USA) in serum samples from LPVM, MPVM and DF patients, and healthy controls following the manufacturer’s instructions. Serum abundances of these five proteins along with three other proteins Ceruloplasmin (CP; P00450), Plasma retinol binding protein (RBP4; P02753), and Plasminogen (PLS; P00747) were measured in the FEB, DEF, and CON stages of the infection and HC following the same assay protocol as described earlier39 (link).
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Proportion of successful displacements from the feed bunk relative to all displacements in which the animal was involved. (Galindo and Broom, 2000) Blood sampling At the end of the experiment, evacuated tubes without anticoagulant were used to collect 10 mL of blood from each animal. Blood samples were centrifuged using 80-2 Electronic Centrifuge (SMIC Inc., China) at 3000 rpm for 15 min. to collect serum. All samples were stored at -20°C until analysis.
Serum triiodothyronine (T3), thyroxine (T4), testosterone and cortisol hormones were estimated by Stat fax-2100 (Awareness technology, INC. USA) and commercial ELISA kits (Assay Max ELISA kits, ASSAYPRO Inc., USA).
Serum total proteins and albumin were assayed by colorimetric method using a commercial kit manufactured by the Egyptian company for biotechnology, Egypt. In addition, serum globulin and Albumin/ Globulin ratio were calculated according to Coles [15] .
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Total serum T4 and T3 were determined in technical duplicates using the total T4 and T3 Coat-a-Count radioimmunoassay kits from Diagnostics Products Corp. (Los Angeles, CA, USA) according to the manufacturer’s instructions. Serum corticosterone was determined using the AssayMax™ ELISA Kit from Assaypro. (St Charles, MO, USA) according to the manufacturer’s instructions.
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To determine treatment group differences in corticosterone, blood from retro-orbital sinus bleeding was centrifuged at 2,000g for 10 min at 4°C, and serum was aliquoted and stored at -80°C. After thawing and diluting samples 1:25 with assay diluent, duplicate serum samples were assayed for corticosterone using an AssayMax ELISA kit following manufacturer’s instructions (AssayPro, St. Charles, MO; minimal detection level: 40 pg/ml; intra-assay and inter-assay coefficients of variation: 5.0 and 7.0%, respectively). Samples with a coefficient variance greater than 15% were repeated. Absorbance was read on a microplate reader at wavelengths of 450 and 570 nm immediately after adding the stopping solution. A wavelength correction was made by subtracting readings at 570 nm from those at 450 nm to correct for optical imperfections. Corticosterone concentrations were determined from standard curves generated from serially diluted standards run in duplicate on each plate.
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Whole blood glycated hemoglobin (HbA1C) (collected in EDTA tubes), and plasma glucose, lipoproteins, and alanine aminotransferase (ALT) were quantified by Cobas c111 analyzer (Roche Diagnostics, Mannheim, Germany). A sensitive rat RIA kit (EMD Millipore, Billerica, MA, USA) was used to measure insulin, and a NEFA-HR2 assay (Wako Chemicals GmbH, Neuss, Germany) was performed to determine non-esterified fatty acids (NEFAs) concentrations. Plasma adiponectin was quantified by AssayMax ELISA kit (Assaypro, St. Charles, MO, USA) and IL1b by Cloud-Clone Corp. (Houston, TX, USA). For liver triglyceride content measurement, 50 mg of frozen liver tissue per sample was weighed on dry ice, and its saponified extract was extracted using a published protocol [18 (link)]. Triglyceride levels were then quantified by Cobas c111 analyzer.
Caffeic acid, trigonelline hydrochloride, cafestol acetate, and ferulic acid were purchased from Sigma-Aldrich (St. Louis, MO, USA) and isotope-labeled hippuric acid (Ring-13C6, 99%) from ReseaLife Chem Science (Burgdorf, Switzerland). HPLC-grade acetonitrile, methanol and acetic acid were purchased from Fluka (Sigma-Aldrich, St. Louis, MO, USA).
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We analyzed the efflux of FABP5 from cells by measuring the amount of FABP5 in the culture supernatant. First, 1.0×10 6 cells were seeded into a 3.5 cm dish, and after a 24-h preincubation, we changed the medium to DMEM without FBS. After a 24-h incubation, the culture medium was collected and centrifuged at 500×g for 10 min to spin down the contaminating cells, and the supernatant was collected for the assay. The collected culture supernatant was concentrated to approximately 1/10 of the initial amount using an ultrafiltration membrane (Amicon Ultra-0.5; Merck, Darmstadt, Germany). FABP5 concentration was measured using a commercial ELISA kit (AssayMax ELISA Kit; Assaypro, St. Charles, MO) according to the manufacturer's protocol. We also measured FABP5 concentration in cell lysates from PLC/PRF/5 and KIM-1 with the total protein concentration adjusted to 100 µg/mL as positive controls and DMEM without FBS as a negative control. All samples were measured in triplicate and analyzed.
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