General Assay Diluent is a mammalian protein-based ELISA additive used to equalize the sample and standard matrices for a more accurate result. Large differences in matrix complexity environment between standard curve and sample wells are the main cause for target analyte under-recovery problems in sandwich ELISA analysis. General Assay Diluent helps equalize matrix complexity disparity issues, reduces non-specific interactions between sample proteins and the plate, and reduces interference from complement and thrombin present in serum and plasma samples.
- 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 General 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.
Boraschi-Diaz I, Tauer JT, El Rifai O, Guillemette D, Lefebvre G, Rauch F, Ferron M, Komarova SV. Metabolic phenotype in the mouse model of osteogenesis imperfecta. J. Endocrinol. 2017 Sep;234(3):279-289. doi: 10.1530/JOE-17-0335. Epub 2017 Jul 17. Abstract.
" – ... 2010b). Briefly, Antibody Coating Buffer (CB1), ELISA Wash Buffer (WB1), 120 General Blocker Buffer (BB1), General Assay Diluent (AD1) and Stop Solution for TMB 121 (STOP1) were all obtained from ImmunoChemistry Technologies. 1-Step™ Ultra TMB ELISA 122 ... "