Prostaglandin E1 ELISA Kit

Sample

Tissue Culture Supernatant, Plasma, Saliva, Serum, Urine

Species Reactivity

Human

Detection Method

cELISA

Intended Use

Prostaglandin E1 ELISA Kit is a competitive immunoassay for the quantitative determination of Prostaglandin E1 in biological fluids.

Contents of Kit

1. Donkey anti-Sheep IgG Microtiter Plate: A plate using break apart strips coated with donkey antibody specific to sheep IgG. 96 Wells
2. PGE1 ELISA Conjugate: A blue solution of alkaline phosphatase conjugated with PGE1. 5 mL
3. PGE1 ELISA Antibody: A yellow solution of a sheep polyclonal antibody to PGE1. 5 mL
4. Assay Buffer: Tris buffered saline, containing proteins and sodium azide as preservative. 27 mL
5. Wash Buffer Concentrate: Tris buffered saline containing detergents. 27 mL
6. Prostaglandin E1 Standard: A solution of 50,000 pg/mL PGE1. 0.5 mL
7. pNpp Substrate: A solution of p-nitrophenyl phosphate in buffer. Ready to use. 20 mL
8. Stop Solution: A solution of trisodium phosphate in water. Keep tightly capped. 5 mL (Caution: Caustic)
9. Plate Sealer: 1 slide

Storage

All components of this kit, except the PEG1 Vonjugate, are stable at 4 °C until the kit's expiration date. The PEG1 Conjugate must be stored at -20 °C.

Precision

Intra-assay precision was determined by taking samples containing low, medium and high concentrations of PGE1 and running these samples multiple times (n=24) in the same assay. Inter-assay precision was determined by measuring three samples with low, medium and high concentrations of PGE1 in multiple assays (n=8). The precision numbers listed below represent the percent coefficient of variation for the concentrations of PGE1 determined in these assays as calculated by a 4 parameter logistic curve fitting program.

Sensitivity

Sensitivity was calculated by determining the average optical density bound for twenty (20) wells run as Bo, and comparing to the average optical density for twenty (20) wells run with Standard #6. The detection limit was the concentration of PGE1, measured at two (2) standard deviations from the zero along the standard curve.
Average Optical Density for the Bo = 0.579 ± 0.012 (2.07%)
Average Optical Density for Standard #6 = 0.558 ± 0.009 (1.61%)
Delta Optical Density (0-4.88 pg/mL) = 0.021
2 SD's of the Zero Standard = 2 x 0.012 = 0.024
Sensitivity = 0.024/0.021 x 4.88 pg/mL = 5.58 pg/mL

General Description

Prostaglandin E1 (PGE1) is synthesized from DGLA, dihomo-γ-linolenic acid. PGE1 has been shown to have a number of biological actions, including vasodilation, proliferation of vascular smooth muscle cells, platelet aggregation and has been shown to have insulin-like actions. Its effects are induced by receptor mediated elevation of cAMP. It is the major prostaglandin in semen.

Standard Curve

The results shown below are for illustration only and should not be used to calculate results from another assay.

A typical standard curve is shown below. This curve must not be used to calculate PGE1 concentrations; each user must run a standard curve for each assay.