Objective We formulated assays for measurement of urinary βLH and βFSH under collection and storage conditions typical of non-clinical research settings. correlated (r = 0.95 for LH 0.86 for FSH). There was no consistent decline with any storage type. Dissociation of subunits by heating was needed for βLH but not βFSH. Conclusion These IEMAs measure free βLH and total βFSH overcoming inter-individual variability in and collection and storage ARRY-438162 effects on subunit dissociation without the need for urine chemical preservatives. Keywords: βLH βFSH IEMA balance Intro Luteinizing hormone (LH) and follicle stimulating hormone (FSH) are delicate signals of hypothalamic-pituitary-ovarian axis function and so are especially valuable for analyzing the timing and correlates of puberty (e.g. (1)) and reproductive ageing (e.g. (2)). A continual issue in the usage of urinary and serum LH and FSH assays can be concern over the consequences of collection and storage space circumstances on dissociation from the alpha and beta (β) subunits of the human hormones (3-6). That is especially relevant for large-scale population-level study where there could be substantial hold off between specimen collection from people within their homes or in the field and period of assay. Many assays for LH and FSH gauge the intact types of the human hormones (7-12) and therefore elements influencing dissociation from the subunits are ever present worries. While limited storage space at refrigerated temps (up to six months) will not appear to influence the balance of undamaged LH and FSH (7 9 13 storage space at freezing or space temperature continues to be found to considerably reduce the quantity of undamaged urinary LH and FSH (7 9 10 13 One study however found no effect of up to 10 freeze-thaw cycles on urinary LH and FSH measurements (12). Although refrigerated temperatures do not appear to influence LH and FSH stability large-scale refrigerator storage is usually impractical in most settings. Preservatives such as glycerol thymol bovine serum albumin boric acid thimerosal and sodium azide or adjustment of specimen pH to neutral are used to prevent subunit dissociation and ensure accurate measurement of intact LH and FSH (e.g. (7 9 10 13 However preservatives have some limitations for population level research including time and expense invested in the advance preparation of specimen collection tubes for home or field collections potential assay interference effects on other hormones to be measured in the same Mouse monoclonal to CD49d.K49 reacts with a-4 integrin chain, which is expressed as a heterodimer with either of b1 (CD29) or b7. The a4b1 integrin (VLA-4) is present on lymphocytes, monocytes, thymocytes, NK cells, dendritic cells, erythroblastic precursor but absent on normal red blood cells, platelets and neutrophils. The a4b1 integrin mediated binding to VCAM-1 (CD106) and the CS-1 region of fibronectin. CD49d is involved in multiple inflammatory responses through the regulation of lymphocyte migration and T cell activation; CD49d also is essential for the differentiation and traffic of hematopoietic stem cells. specimen and the fact that dilution volume correction may be necessary for the final urinary result (3 ARRY-438162 13 Additionally preservatives do not correct for any inter-subject variability that might exist in modification of LH and FSH between circulation in the blood and excretion in the urine (3). Our objective was to develop immunoenzymometric assays to measure LH and FSH in spot urines for population-level research that are robust to non-clinical collection and storage of urine specimens and do not require specimen preservation or extraction. Following the approach of Qiu et al. (3) we identified antibodies specific to βFSH and βLH and developed assays that would measure the total amount of each β subunit in urine specimens whether they were in intact or dissociated form. METHODS Samples A total of 799 daily urine and serum specimens were collected over one menstrual cycle from 30 US women in 1997-1998. Thirteen women aged 20-25 years and 17 women 40-45 years old were recruited for a study on reproductive aging. Monetary compensation was provided participants provided written informed consent and all procedures were approved by the Institutional Review Board of the University of Washington. All participants were normally cycling in good health had a ARRY-438162 mean body mass index of 22.6 kg/m2 (SD = 2.36 range 18.9 – 27.7) and were not using medications or hormones. Blood specimens were obtained by venipuncture beginning with the first day of menstrual bleeding and continuing until the first day of menstrual bleeding of the subsequent cycle. Serum specimens were assayed within 1 to 2 2 months of collection for intact LH and FSH. The LH assay (DELFIA Wallac Inc. Gaithersburg MD USA) cross-reacts less than 1% with FSH and the inter- and intra-assay CVs were 2.8% and 4.7% respectively. The FSH assay (DELFIA Wallac Inc. Gaithersburg MD USA) cross reacts less than 1% with LH and the inter- and intra-assay CVs were 2.3% and 4.6% respectively. All cycles were confirmed ovulatory by transvaginal ultrasound. Urine specimens were taken ARRY-438162 daily in the center before noon at exactly the same time seeing that usually.