I am trying to find out the appropriate ELISA method to detect aflatoxin-lys from serum sample. I am facing problems with sample preparation before ELISA. Currently I am following the steps: 1. Isolation of alb from serum sample by ammonium sulphate 2. quantitation of alb by comassie protein assay 3. digestion of alb by pronase to get af-lys 4. ELISA to quantify the af-lys using AFB-lys as standard. But its not working well because I am having a lower level of alb from serum sample than expected or it may be other reasons. I would appreciate any help you all may be able to give me. Thanks in advance.
bonjour,
ce que j'ai compris voulez vous faire dosage de l'aflatoxine dans le serum, et vous voulez déprotieniser, mais si votre molecule est lié au protein (albumine),
dans cette technique (ELISA) le meilleur est d'utiliser des anticorps monoclonaux,
d'autre part je pense vous devez quantifier l'aflatoxine dans le serum avant et après déproteinisation c à d (fair le dosage dans le serum entire, le surnageant de la déproteinisation et dans la partie proteique après digestion)
You could use ready to use aflatoxin kits by Randox. These kits are approved from multiple matrices including serum and meat.
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