Pregnanediol-3-Glucuronide (PDG) ELISA Kit

Name: Pregnanediol-3-Glucuronide (PDG) ELISA Kit
SKU: DEIABL255
Rating: 5 (1 reviews)

Regulatory status: For research use only, not for use in diagnostic procedures.

COA

SDS

Datasheet

Summary

Sample

Dried Fecal Extracts, Urine, Extracted Serum/Plasma, and Tissue Culture Media

Detection Method

cELISA

Intended Use

The Pregnanediol-3-Glucuronide (PDG) Immunoassay kit uses a specifically generated antibody to measure PDG and its metabolites in urine and fecal samples, or in extracted serum and plasma. This kit is not recommended for serum, plasma, or saliva samples without extraction. The kit will quantitatively measure PDG present in diluted buffer samples and tissue culture media samples.

Contents of Kit

1. Coated Clear 96 Well Plates
Clear 1 by 8 break-apart strip well plastic microtiter plate(s) coated with goat anti-rabbit IgG.
2. Pregnanediol-3-Glucuronide (PDG) Standard
Pregnanediol-3-Glucuronide (PDG) at 500 ng/mL in a special stabilizing solution. 125 μL
3. Pregnanediol-3-Glucuronide (PDG) Antibody
A rabbit polyclonal antibody speciffc for Pregnanediol-3-Glucuronide. 3 mL
4. Pregnanediol-3-Glucuronide (PDG) Conjugate
Pregnanediol-3-Glucuronide-peroxidase conjugate in a special stabilizing solution. 3 mL
5. Assay Buffer Concentrate
A 5× concentrate that should be diluted with deionized or distilled water. 28 mL
6. Wash Buffer Concentrate
A 20× concentrate that should be diluted with deionized or distilled water. 30 mL
7. TMB Substrate. 11 mL
8. Stop Solution
A 1M solution of hydrochloric acid. CAUSTIC. 5 mL
9. Plate Sealer

Storage

All components of this kit should be stored at 4°C until the expiration date of the kit.

Precision

Intra Assay Precision
Three urine samples were diluted with Assay Buffer and run in replicates of 20 in an assay. The mean and precision of the calculated pregnanediol-3-glucuronide (PDG) concentrations were:

Inter Assay Precision
Three urine samples were diluted with Assay Buffer and run in duplicates in fourteen assays run over multiple days by three operators. The mean and precision of the calculated pregnanediol-3-glucuronide (PDG) concentrations were:

Sensitivity

Sensitivity was calculated by comparing the OD's for twenty wells run for each of the B0 and standard #8. The detection limit was determined at two (2) standard deviations from the B0 along the standard curve. Sensitivity was determined as 0.180 ng/mL.
The Limit of Detection for the assay was determined in a similar manner by comparing the OD's for twenty runs for each of the zero standard and a low concentration human urine sample. Limit of Detection was determined as 0.320 ng/mL

General Description

Pregnanediol Glucuronide, C₇H₄₄O₈, also known as PDG (5β-Pregnan-3a,20a-diol 3-glucosiduronate) is the major metabolite of progesterone^1-4. Progesterone is the hormone involved in the female menstrual cycle, gestation and embryogenesis of humans and other species. Progesterone belongs to a class of hormones called progestogens, and is the major naturally occurring human progestogen^5,6. Progesterone is an essential regulator of human female reproductive function in the uterus, ovary, mammary gland and brain, and plays an important role in non-reproductive tissues such as the cardiovascular system, bone and the central nervous system. Progesterone action is conveyed by two isoforms of the nuclear progesterone receptor (PR), PRA and PRB. PRA and B are expressed in a variety of normal breast tissue from humans, rats and mice and is also expressed in breast cancer cells^7,8. Progesterone also has neurotrophic roles in the peripheral nervous system as it activates the growth and maturation of axons and stimulates the repair and replacement of myelin sheaths in regenerating nerve fibres⁹.

1. Tait, J.F., Little, B., Tait, S.A.S. et al. Excerpta Medica (Interim Congress Series), 1962, 51:13.
2. Arcos, M., Gurpide, E., Vande Wiele, R.L. and Lieberman, S. Precursors of urinary pregnanediol and their inffuence on the determination of the secretory production rate of progesterone. J. Clin. Endocrinol., 1964, 24:237–245.
3. Lasley, B.L., Stabenfeldt, G.H., Overstreet, J.W. et al. Urinary hormone levels at the time of ovulation and implantation. Fertil. Steril., 1985, 43:861–867.
4. Stanczyk, F.Z., Miyakawa, I. and Goebelsmann, U. Direct radioimmunoassay of urinary estrogen and pregnanediol glucuronides during the menstrual cycle. Am J Obstet Gynecol, 1980, 137:443-450.
5. Collins, W.P., Branch, C.M., Collins, P.O. and Sallam H.N. Biochemical indices of the fertile period in women. Int J Fertil., 1981, 26:196-202.
6. Brown, J.8. and Gronow, M. Endocrinology of ovulation prediction, in Clinical Reproductive Endocrinology (R.P. Shearman, Editor). Churchill Livingstone, Edinburgh, 1985, p. 165-184.
7. Wiebe JP. Progesterone metabolites in breast cancer. Endocr Relat Cancer 2006, 13(3):717–738.
8. Lukanova A, Kaaks R. Endogenous hormones and ovarian cancer: epidemiology and current hypotheses. Cancer Epidemiol Biomarkers Prev 2005;14:98–107.
9. Koenig, HL, Gong, WH and Pelissier, P., "Role of progesterone in peripheral nerve repair." Revs. of Reprod., 2000; 5:189-199.

Standard Curve

Average the duplicate OD readings for each standard and sample. Create a standard curve by reducing the data using the 4PLC fitting routine on the plate reader, after subtracting the mean OD's for the NSB. The sample concentrations obtained, calculated from the %B/B0 curve, should be multiplied by the dilution factor to obtain neat sample values.