Wetlab

Click here to take a tour of the Wetlab.

Methods

Our laboratory assays include urinary enzyme immunoassays for five of the primary hormones regulating female reproduction.

We have assays for estrone-3-glucuronide (the main urinary metabolite of estradiol), preganediol-3-glucuronide (the main urinary metabolite of progesterone), luteinizing hormone (LH), follicle-stimulating hormone (FSH), and human chorionic gonadotropin (hCG, the pregnancy hormone).

Our steroid hormone assays are competitive EIA's, following the format of Munro et al. (1991). Our protein hormone assays are sandwich ELISA's, and follow the formats of Holman et al. (1998) (for intact hCG) O'Connor et al. (for intact, beta and beta core hCG) and Qui et al. 1998 (for LH and FSH).

Protocols for our E1C, E1G, PDG, FSH, hCG and LH enzyme immunoassays are available upon request. Please contact Eleanor Brindle at ebrindle@u.washington.edu.

The lab is fully equipped for large scale processing of all of these assays. For example, we use a robotic pipettor (fondly, or not so fondly, called Hal; see wetlab tour above) to help us process up to 2400 samples a week.

References cited:

O’Connor KA., Brindle E, Shofer JB, Miller RC, Klein NA, Soules MR, Campbell KL, Mar C, Handcock MS (2004). Statistical correction for non-parallelism in a urinary enzyme immunoassay. Journal of Immunoassay and Immunochemistry 25 (3): 259-278.

Miller RC, Brindle E, Holman DJ, Shofer J, Klein NA, Soules MR, O’Connor KA. (2004). Comparison of specific gravity and creatinine methods for normalizing urinary reproductive hormone concentrations. Clinical Chemistry 50: 924-932.

O’Connor KA, Brindle E, Holman DJ, Klein NA, Soules MR, Campbell KL,
Kohen F, Munro CJ, Shofer J, Lasley WL, Wood JW. (2003) Urinary estrone conjugate and pregnanediol-3-glucuronide enzyme-immunoassays for population research. Clinical Chemistry 49(7):1139-1148.

Holman DJ, Rasheed FN, Stroud CM, Brindle E, O'Connor KA, and Campbell KL (1998) A commercial pregnancy test modified for field studies of fetal loss. Clinica Chimica Acta 271:25-44.

O'Connor JF, Schlatterer JP, Birken S, Krichevsky A, Armstrong EG, McMahon D, Canfield RE (1988). Development of highly sensitive immunoassays to measure human chorionic gonadotropin, its beta-subunit and beta core fragment in the urine: Application to malignancies. Cancer Research 48:1361-1366.

Qiu Q, Kuo A, Todd H, Dias JA, Gould JE, Overstreet JW, Lasley BL (1998) Enzyme immunoassay method for total urinary follicle-stimulating hormone (FSH) beta subunit and its application for measurement of total urinary FSH. Fertiity andl Sterility 69(2):278-85.