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The Microwell Progesterone ELISA is an enzyme immunoassay system for quantitative determination of progesterone levels in serum/plasma.

Rodent Progesterone ELISA Kit

Background

Progesterone is a steroid hormone (C21 steroid, pregn-4-ene-3, 20 dione) and is synthesized from both tissue and circulating cholesterol. The principal production sites are the adrenals and ovaries and placenta during pregnancy. The majority of this steroid is metabolized in the liver to pregnanediol and conjugated as a glucuronide prior to excretion by kidneys. The primary role is played in reproductive organs. In males the progesterone plays an intermediary role in the synthesis of corticosteroids and androgens. In females, progesterone remains relatively constant through follicular phase of ovulation. Then the levels increase following ovulation and remains elevated for several hours and decreasing to the base line 24 hours before the onset of cycle. In pregnancy, placental progesterone raise 10-30 times those of the luteal peak levels.
In female rodents, the measurement of progesterone is an useful marker in evaluating the status of ovarian functions and also pregnancy.

Test Principle

The progesterone quantitative Test is based on a solid-phase enzyme immunoassay based on competitive binding method. A sample (serum/ plasma/urine) containing an unknown amount of progesterone will compete with enzyme-conjugated progesterone for high affinity binding sites on a limited number of antibodies coated on to the plate. After washing away the free antigen, the amount of labeled antigen in the sample is reversibly proportional to the concentration of the unlabeled antigen. The actual concentrations in unknown samples are obtained by means of a standard curve based on known concentrations of unlabeled antigen analyzed in parallel with the unknowns. After washing, substrate solution is added and the enzyme allowed to react for a fixed time before the reaction is terminated. Absorbencies are measured at 450 nm using ELISA plate reader. A standard curve is produced using values from standards from which absorbency values for blank tubes have been subtracted. Results for unknown may be read directly from this standard curve using either manual calculation or by a suitable computer program. This kit is suitable for the direct measurement of progesterone in serum/plasma samples. It may also be used following.

Feature and Advantages

<Quality Control Data>
A typical standard curve (illustration only) for rodent Progesterone is given below:

Standard ng/mL OD at 450nm
10 0.05
5 0.24
2.5 0.64
1 0.96
0.5 1.5
0 2.6

<ELISA Performance Characters>
Precision : Inter and Intra assay variation (CV) was determined from three different pooled serum samples in three differentexperiments.

Inter-assay variation Set1: CV= 5.8 % (N=10) Set2: CV= 5.6 % (N=10) Set3: CV= 4.9 % (N=10)
Intra-assay variation Set1: CV= 4.6 % (N=10) Set2: CV= 5.2 % (N=10) Set3: CV= 6.2 % (N=10)

< Sensitivity >
The lowest level detectable in this assay is 0.1ng/mL of serum or plasma

< Specificity >
The rodent Progesterone ELISA system utilizes Highly specific Progesterone antibody Coated on to the plate. The cross-reactivity to other related hormones is not detectable under the sensitivity of the assay system.

< References >
1. Kasson BG. et al. 1993 Characterization of a rapid and sensitive enzyme immunoassay (EIA) for progesterone applied to conditioned cell culture media. J Immunoassay 14 33-49
2. Roby KF. 2001 Alterations and follicle development, steroidogenesis and gonadotropin receptor binding in a model of ovulatory blockade. Endocrinology 142 2328
3. Vaitukaitis J. Robbins JB. Nieshlag E. Ross GT J. Clin. Endocrinol. Metab. 33 988-991. (1971).
4. Nakane PK. and Kawaoi A. J. Histochem. Cytochem. 22:1084-1091(1974).
5. Maiolini R. and Masseyeff R. J. Immunol. Methods. 8:223-234 (1975).
6. Hsu, YH. Anal. Biochem. 142:221-225, (1984).
7. Chattoraj SC 1976 Endocrine function in Fundamentals of Clinical Chemistry, NW Tietz eds WB Saunders Chap 13, 699-82

Kit component

1. Wells coated with progesterone antibody (96 wells)
2. Enzyme Conjugate, 12 mL
3. Sample/standard diluent 25mL
4. TMB Color Reagent, 12 mL
5. Stop Solution (2N HCL), 6 mL.
6. 20 X Wash Buffer, 20mL.
7. Progesterone Standard Set: 0, 1.0, 2.5, 5.0, 10, 30 ng/mL 0.6 ml /Vial
QC1 ( ~2.5 ng/mL and QC2 , 10 ng/mL) 0.6mL/Vial
8. Instruction

Product List

Product Name Cat# Quantity Price

Progesterone ELISA

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Estradiol ELISA

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€ 1250