- Once the capture antigen has been coated onto the plate and the plate has been properly blocked with one of ICT’s stabilizing blocker products, pipette 50 – 100 µL of IgM-Reducing Assay Diluent into each of the wells of the ELISA plate using a multi-channel pipettor. The addition of the Assay Diluent is always done prior to adding the standard or test samples to each well.
- The standard and test samples should be diluted to the proper well concentrations using one of ICT’s three different Sample Diluent products to help further equalize matrix complexity parameters between standard/calibrator wells and the serum/plasma containing sample wells.
- Because IgM-Reducing Assay Diluent contains a more diverse composition of additives, it can have some negative impact on the antigen capture step of the ELISA. This leads to reduction in ELISA sensitivity which must be compensated for by using longer antigen capture incubation times or more sensitive HRP substrate components.
Niu H, Klem T, Yang J, Qiu Y, Pan L. A biotin-drug extraction and acid dissociation (BEAD) procedure to eliminate matrix and drug interference in a protein complex anti-drug antibody (ADA) isotype specific assay. J Immunol Methods. 2017. Jul;446:30-36. Epub 2017 Apr 5. Abstract.
" – ... Sulfo-tag™-conjugated anti-human/NHP IgG or IgM Ab was purchased from MSD (Meso Scale Discovery, Rockville, Maryland). IgM-reducing assay diluent was purchased from ImmunoChemistry Technologies (Bloomington, MN). ... "