Table 2.

Preparation of soluble substances

Source material	Preparation
Guinea pig urine	After collection of urine, boil for 10 minutes. Dialyze for 48 h at 4°C against multiple changes of pH 7.4 PBS. Centrifuge, aliquot, and freeze.19
Human milk	Collect milk from lactating women, centrifuge at 1000g (serofuge at 3400 rpm) for 10 minutes, remove and discard the cream layer, incubate milk in boiling water for 10 minutes, mix 1 volume of milk with 1 volume of PBS. Aliquot and freeze.2
Human saliva	Collect 2 mL saliva, boil for 10 minutes, centrifuge at 1000g (serofuge at 3400 rpm) for 10 minutes, harvest supernate, aliquot, and freeze.2
Human urine	Collect urine from three individuals, pool, centrifuge, dilute with equal volume of distilled water, check pH (dialysis with PBS may be needed to obtain pH between 6 and 8.5), aliquot, and store frozen until needed.2
Hydatid cyst fluid	Incubate HCF (with scolices) from animal or human sources at 56°C for 1 hour. Dilute 1 volume of hydatid cyst fluid with 9 volumes of PBS. Aliquot and freeze.2
Pigeon egg albumin	Separate egg white from the yolk. Prepare dilutions of 1:100 to 1:1000 in PBS. Test the dilutions using a potent anti-P1 to determine the best dilution for inhibition studies. Make aliquots of appropriate dilution and freeze.2
Pooled plasma	Purchase commercial pooled plasma or prepare pooled plasma from six or more donors. Aliquot and freeze.3
Recombinant protein	May be commercially available.5–7,15

PBS = phosphate-buffered saline.