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The Microwell Estradiol ELISA is an enzyme immunoassay system for quantitative determination of Estradiol levels in rodent serum.

Rodent Estradiol ELISA Kit

Background

The E2 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 E2 to be assayed (unlabeled antigen) is added to a standard amount of a conjugated E2(labeled antigen). The labeled and unlabeled antigens are then allowed to compete for high affinity binding sites of E2 antibody on a limited number of secondary 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 is 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 5 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 E2 in serum samples.

Feature and Advantages

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

Standard pg/mL OD at 450nm
5000 0.35
1200 0.54
300 0.81
100 0.98
30 1.4
10 1.6
0 1.99

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

Inter-assay variation Set1: CV= 6.8 % (N=10) Set2: CV= 7.9 % (N=10) Set3: CV= 7.8 % (N=10)
Intra-assay variation Set1: CV= 6.9% (N=10) Set2: CV= 8.9 % (N=10) Set3: CV= 8.6 % (N=10)

< Sensitivity >
The lowest level detectable in this assay is 5 pg/mL of serum or plasma

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

< References >
1. Edwards RG, et.al.” Lancet 11:611, 1972.
2. Larsen S. Honore E: .”Feril Steril. 30:745, 1978.
3. Muechler EK, et.al Fertil. Steril. 30:745, 1978.
4. Shaaban MM & Klopper A “ A study on the monitoring of gonadotropin therapy by the assay of plasma E2 and Progesterone” J Obstet Gynecol Br.Commonw 80 783, 1973
5. Smith DH et al. Fertil. Steril 33 387 1980
6. Vaitukaitis J. Robbins JB. Nieshlag E. Ross GT J. Clin. Endocrinol. Metab. 33 988-991. (1971).
7. Nakane PK. and Kawaoi A. J. Histochem. Cytochem. 22:1084-1091(1974).
8. Maiolini R. and Masseyeff R. J. Immunol. Methods. 8:223-234 (1975).
9. Hsu, YH. Anal. Biochem. 142:221-225, (1984).
10. Chattoraj SC 1976 Endocrine function in Fundamentals of Clinical Chemistry, NW Tietz eds WB Saunders Chap 13, 699-823

Kit component

1. Microtiter wells 96, coated with second antibody.
2. Rabbit anti E2 antibody 7.0 mL
3. Enzyme Conjugate solution, 12 mL.
4. TMB Color Reagent, 12 mL
5. Stopping Solution (2N HCL), 6 mL
6. 20 X Wash Buffer, 20 mL.
7. Sample diluent, 20 mL
8. E2 Standard set: 0,10, 30,100, 300, 1200, 5000pg/mL.(+ QC1 & QC2)
9. Instructions

Product List

Product Name Cat# Quantity Price

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