Detection and titration of specific anti-AFB1 Abs were carried out by ELISA, as previously described [15] (link). Briefly, serially ten-fold diluted (from 1∶4 and 1∶10 to 1∶400,000) immune serum or control (pre-immune serum) was added to each well of polystyrene microtiter plates coated with AFB1-BSA conjugate (Sigma-Aldrich) or BSA control protein. After incubation, reaction was detected by adding rabbit anti-bovine IgG (whole molecule) peroxidase conjugate Abs (Sigma-Aldrich, product number A7414) and chromogen/substrate solution. The optical density (OD) at 450 nm was read by using a Multiskan Ascent spectrophotometer (Labsystems, Helsinki, Finland), and the titer of each immune serum was defined as the inverse of the highest dilution that gave 0.1 OD above the pre-immune serum at the same dilution. To compensate for between-plate variability, individual plates were normalized to the mean of the appropriate positive control. For statistic evaluation the logarithms of anti-AFB1 Ab titers were considered.
Afb1 bsa conjugate
Manufactured by Merck Group
AFB1-BSA conjugate is a laboratory product that consists of aflatoxin B1 (AFB1) chemically conjugated to bovine serum albumin (BSA). This conjugate is used as a reference standard or antigen in various analytical and immunological applications, such as the development and validation of detection methods for aflatoxin B1.
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Lab products found in correlation
2 protocols using afb1 bsa conjugate
1
Enzyme-Linked Immunosorbent Assay for Anti-AFB1 Antibodies
2
Quantitative Immunoassay Reagent Preparation
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All reagents were of analytical grade. 35.7 mM of 3,3′,5,5′-tetramethylbenzidine chromogen reagent (Dojindo Laboratories) was prepared with acetonitrile. 1× blocker bovine serum albumin in phosphate-buffered saline (BSA-PBS) (Thermo Fisher Scientific Inc.) was diluted with 1× phosphate-buffered saline Tween® 20 (PBST), pH 7.5 (Thermo Fisher Scientific Inc.), which was used as the washing solution during immunoassay. For the biotin immunoassay, 200 μg mL -1 biotin stock solution (Sigma-Aldrich, Inc.) and 10 mg mL -1 biotin-BSA conjugate (Sigma-Aldrich, Inc.) were prepared by dissolving the solids separately in PBST. For the aflatoxin B 1 (AFB 1 ) immunoassay, 1 mg AFB 1 powder (Sigma-Aldrich, Inc.) was dissolved in 1 mL acetonitrile, and 1 mg mL -1 AFB 1 -BSA conjugate (Sigma-Aldrich, Inc.) was dissolved in PBST.
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