Affinity and Matrix Effects in Measuring Fish Plasma Vitellogenin Using Immunosorbent Assays: Considerations for Aquatic Toxicologists
Figure 2
Standard curves generated using polyclonal antisera (a) or monoclonal (b) antibodies against fathead minnow vitellogenin. Standard vitellogenin was diluted across seven points in three different matrices. Normal method: prepared in phosphate-based dilution buffer. Diluted plasma: unexposed plasma substituted for dilution buffer, where the plasma was diluted with the standards across a range of 1 : 16 to 1 : 1024. Constant plasma: unexposed plasma substituted for dilution buffer with standards added, where the plasma was diluted 1 : 500. Unexposed fathead minnow plasma confirmed by ELISA for undetectable vitellogenin at a detection limit of 3 μg/mL.
