The measured values of urinary PTGDS fell on the linear portion of the ELISA kit standard curve.
The specific activity of IFN-γ kit standard was titrated against NIBSC IFN-γ European standard and was set at 3.61 IU for one ng IFN-γ.
Briefly, lyophilised Quantikine standard was reconstituted in distilled water and serially diluted one in two in kit standard diluent to produce a seven-point standard curve ranging from 15.6 pg/ml to 1000 pg/ml.
Approximately 1 µL of purified peptides and 1 µL of ProteoMass Peptide & Protein MALDI-MS Calibration Kit standard (Sigma) were spotted onto a MTP 384 Massive MALDI target (Bruker Daltonics, Billerica, MA).
Then sections were incubated in ABC solution (ABC kit standard PK-4000; Vector LaBurlingame, Burlingame, CA) for 90 min at room temperature and peroxidase activity revealed for 5 10 min with nickel-enhanced DAB (DAB-Ni), resulting in a black precipitate.
In order to assess recovery, supernatant samples SN1, SN2 and SN3 were each spiked at three concentrations (ranging from 7 5461 pg/ml) with recombinant cytokine using the Bio-plex kit standard.
Total genomic DNA was isolated from saliva samples using Oragene-DNA Self-Collecting kit standard protocols.
The kit standard curve is validated to measure suPAR levels between 0.6 to 22.0 ng/ml with a detection limit (LLOD) of 0.1 ng/ml.
The SD BIOLINE Malaria Ag P.f/P.v POCT test kit (Standard diagnostic, Inc, Germany, Lot No. 145021) was used to capture malaria antigens and was performed in accordance with the manufacturer's instructions.
Samples were split into two aliquots and one aliquot was run as "neat or unspiked", while the other aliquot was "spiked" with a small volume of the kit standard concentrate, then compared to a "spiked" diluent.
IgG1 and IgG4 standard curves were calibrated against a set of PeliClass human IgG subclass ELISA kit standards (WHO 67/97 reference preparation) (Cell Sciences, Canton, MA, USA).
