Aflatoxin b1 bovine serum albumin afb1 bsa

The total accumulation of aflatoxins was estimated for both control and infected seeds following the protocol suggested by Waliyar et al. [14 ] using indirect competitive enzyme-linked immunosorbent assay (ELISA). In this, we used polyclonal antibodies that were produced against aflatoxin B₁ (AFB₁) for quantitative estimation of total aflatoxins. Aflatoxin B1-Bovine Serum Albumin (AFB₁-BSA), obtained from Sigma (Catalogue No. 6655, Suffolk, NY, USA), was used for the production of polyclonal antibodies to AFB₁. Polyclonal antibodies used in the indirect competitive ELISA were produced in New Zealand White inbred rabbit following the protocol developed in our institute previously [15 (link)]. The seed coats of the control and infected genotypes were stained with Coomassie Brilliant Blue and were observed through a Florescence microscope (Zeiss Axio Scope.A1, Carl Zeiss, Oberkochen, Germany) to visualize fungal colonization and sclerotia formation.

Quantitative estimation was done for total aflatoxins under control and infected conditions following the protocol of Waliyar et al. [36 ]. An indirect competitive enzyme-linked immune sorbent assay (ELISA) approach was used, and polyclonal antibodies were produced against AFB₁) [36 ]. To do so, we utilized polyclonal antibodies against AFB₁ for the quantitative assessment of all of the aflatoxins. For polyclonal antibody production, aflatoxin B₁-bovine serum albumin (AFB₁-BSA) was acquired from Sigma-Aldrich (Catalog No. 6655, Suffolk, NY, USA).