The reactivity of polyclonal antibodies from infected or non-infected animals was evaluated by ELISA. Briefly, ninety-six-well microtiter plates (Nunc, Roskilde, Denmark) were coated overnight at 4°C with 1 μg/well of FhTE in 50 mM carbonate buffer (pH 9.6). After blocking with 1% BSA in PBS, three washes with PBS containing 0.1% Tween-20 were performed. For the oxidation of the glycan moieties of FhTE, wells were treated with 10 mM of sodium meta-periodate in 50 mM sodium acetate buffer pH 4.5 for 30 min at room temperature in the dark, washed with 50 mM sodium acetate buffer and subsequently incubated for 1 h with glycine 1% at room temperature. As controls (Fhmock), wells were subjected to the same treatment except for the incubation with sodium meta-periodate. Serially diluted sera in buffer (PBS containing 0.1% Tween-20 and 0.5% BSA) were added to the wells for 1 h at 37°C. Following three washes, wells were treated 1 h at 37°C using goat anti-mouse polyvalent peroxidase-conjugate (Sigma-Aldrich, St. Louis, MO) and o-phenylenediamine-H2O2 was then added as substrate. Plates were read photometrically at 492 nm in an ELISA auto-reader (Labsystems Multiskan MS, Finland).
Ninety six well microtiter plates
Manufactured by Thermo Fisher Scientific
Sourced in United States
Ninety-six-well microtiter plates are a type of laboratory equipment used for various applications in life science research and diagnostics. These plates have a grid of 96 individual wells that can hold small volumes of liquid samples, typically in the range of microliters. The primary function of these plates is to facilitate the processing, storage, and analysis of multiple samples simultaneously in a standardized and high-throughput manner.
Automatically generated - may contain errors
Lab products found in correlation
1 ethyl 3 3 dimethylaminopropyl carbodiimide hydrochloride edc, by Dojindo Laboratories (1 mentions)
Horseradish peroxidase hrp, by Toyobo (1 mentions)
Goat anti mouse igg h l antibody, by Thermo Fisher Scientific (1 mentions)
N hydroxysuccinimide nhs, by Fujifilm (1 mentions)
Nunc maxisorp, by Thermo Fisher Scientific (1 mentions)
2 protocols using ninety six well microtiter plates
1
Evaluating Polyclonal Antibody Reactivity by ELISA
2
Mycotoxin Analysis Protocol Optimization
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AFB 1 , AFB 2 , AFG 1 and AFG 2 , which were of analytical grade for mycotoxin analysis, and N-hydroxysuccinimide (NHS) were purchased from Wako Pure Chemical Industries, Ltd. (Osaka, Japan). 1-Ethyl-3-(3-dimethyl- aminopropyl)carbodiimide hydrochloride (EDC) was purchased from Dojindo Molecular Technologies, Inc.
(Kumamoto, Japan). Bovine serum albumin (BSA; product no. A7888) was purchased from Sigma-Aldrich Co. LLC (St. Louis, MO, USA). Ninety-six-well microtiter plates (Nunc MaxiSorp) and goat anti-mouse IgG (H＋L) antibody (Pierce) were obtained from Thermo Fisher Scientific (Waltham, MA, USA). Horseradish peroxidase (HRP) was purchased from Toyobo Co., Ltd. (Osaka, Japan).
Roasted peanuts produced in Kanagawa prefecture, Japan, were obtained from the market, and it was confirmed by LC-MS/MS that they were not contaminated with AFs. Ground food samples contaminated with AFB 1 , AFB 2 , AFG 1 and AFG 2 were provided by the Food Analysis Performance Assessment Scheme (FAPAS): peanuts (T04118), almond (T04114), pistachio (T04283) and chili powder (T04286). Commercial peanut butter from the USA (17 samples), India (2 samples), or Japan (4 samples) was purchased from markets in the Kansai area of Japan.
All other chemicals and reagents, which were of analytical grade, were purchased from Wako Pure Chemical Industries, Ltd. or Nacalai Tesque, Inc. (Kyoto, Japan).
(Kumamoto, Japan). Bovine serum albumin (BSA; product no. A7888) was purchased from Sigma-Aldrich Co. LLC (St. Louis, MO, USA). Ninety-six-well microtiter plates (Nunc MaxiSorp) and goat anti-mouse IgG (H＋L) antibody (Pierce) were obtained from Thermo Fisher Scientific (Waltham, MA, USA). Horseradish peroxidase (HRP) was purchased from Toyobo Co., Ltd. (Osaka, Japan).
Roasted peanuts produced in Kanagawa prefecture, Japan, were obtained from the market, and it was confirmed by LC-MS/MS that they were not contaminated with AFs. Ground food samples contaminated with AFB 1 , AFB 2 , AFG 1 and AFG 2 were provided by the Food Analysis Performance Assessment Scheme (FAPAS): peanuts (T04118), almond (T04114), pistachio (T04283) and chili powder (T04286). Commercial peanut butter from the USA (17 samples), India (2 samples), or Japan (4 samples) was purchased from markets in the Kansai area of Japan.
All other chemicals and reagents, which were of analytical grade, were purchased from Wako Pure Chemical Industries, Ltd. or Nacalai Tesque, Inc. (Kyoto, Japan).
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