Quantitative test is based on the principle of 2. Dispense 10 uL of standards, controls or competes with enzyme-labeled-Progesterone 3. Progesterone Antibody Solution (6 mL) Progesterone
for a fixed and limited number of antibody 3. Dispense 50 uL of antibody solution into In the assay procedure, the Progesterone standard or patient serum is incubated with NAME AND INTENDED USE
conjugate in the anti-mouse IgG coated well. In this solid-phase system, the antibody bound Progesterone will remain on the well Quantitative SH-301 is a solid phase enzyme while unbound Progesterone will be removed linked immunosorbent assay (ELISA). This TMB substrate, is mixed with the antibody 7. TMB Solution (11 mL): Buffer solution test provides quantitative measurement of bound Progesterone-horseradish peroxidase enzyme conjugate. After a short incubation, 9. Well holder for securing individual wells. microreader at 450 nm. When high levels of SUMMARY AND EXPLANATION OF TEST
enzyme conjugate is bound; then, less color PROVIDED
Progesterone (Pregn-4-ene-3, 20-dione) is a C21 steroid hormone. The molecular weight 2. Pipetor with tip for 10 uL, 50 uL and PROCEDURAL NOTE
Progesterone is a female sex hormone that WARNING AND PRECAUTION
in conjunction with estrogens, regulates the 1. Azide and thimerosal at concentration accessory organs during the menstrual cycle higher than 0.01% interfere in this test; and it is particularly important in preparing Quantitative is designed for in vitro use SPECIMEN COLLECTION AND HANDLING
the endometrium for the implantation of the blastocyte and in maintaining pregnancy. In 2. The components in this kit are intended Collect blood by venipuncture, allow clotting, non-pregnant women progesterone is mainly and separating the serum by centrifugation secreted by the corpus luteum whereas in 3. The components of different lots should pregnancy the placenta becomes the major assayed immediately, they can be stored at exchange strips of different plates even 2 - 8oC or frozen for few days. Do not use cortex for both sexes and the testes for hyperlipemic, hemolyzed, contaminated or 3. Pippet all reagents and samples into the heat inactivated sample as they may cause conditions, and the characteristics of the bound to Corticosteroid Binding globulin 4. Absorbance is a function of the time and (CBG), Sex Hormone Binding globulin (SHBG) PREPARATION FOR ASSAY
concentration circulates as free hormone. Store the kits at 2-8oC in the refrigerator. Keep microwells in dry bag with desiccants. are clinically used to confirm ovulation and The reagents are stable until expiration of 2. Before the start of the assay. Once the normal function of the corpus luteum in non- the kit. The calibrators and conjugate are stable for 8 weeks after first opening. TMB without any interruption to get the most substrate solution should be colorless; if the Abnormal progesterone secretion has been solution turns blue, it must be replaced. Do implicated in premenstrual tension, irregular not expose these reagents to strong light MATERIALS PROVIDED
1. Microwell strips (96 wells): anti-mouse standards, controls and patient samples. REFERENCES
control at several levels to monitor assay performance. The control should be treated PRECISION
as unknown. Results obtained should be in agreement with the assigned value of the Intra-assay and Inter-assay coefficient of involving the biotin-streptavidin system reference standard against it’s A/Ao x control. The controls can be obtained from variation were evaluated at three different commercially available sources, but should not contain sodium azide as preservatives. Intra - assay
3. Elder P.A., K.H.J. Yeo, J.G. Lewis, and variable results samples containing > 40 ng/mL Progesterone can be diluted with the Inter - assay
A/Ao x 100
containing <0.4 ng/mL are analyzed by Controls
calibrator with the 0 calibrator to extend the ACCURACY
Recovery studies were performed by mixing an aliquot of pooled serum and progesterone clinical finding or impressions, clinical judgment should be exercised and additional EXPECTED VALUE
1. It is recommended that each laboratory SPECIFICITY
The Concentration
of Progesterone

Source: http://www.unitedbioresearchinc.com/elisa/pdf/SH301.pdf


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