Endothelin-1, BioAssay ELISA Kit (Human)

£1,406.00 1 Kit

Endothelin-1, Human BioAssay? ELISA Kit is a 4.5 hour solid phase ELISA designed to measure ET-1 in cell culture supernates, serum, plasma, and urine. It contains synthetic ET-1 and antibodies raised against synthetic ET-1. This immunoassay has been shown to accurately quantitate synthetic and naturally occurring ET-1.

Principle of the Test:
This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal antibody specific for ET-1 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any ET-1 present is bound by the immobilized antibody. After washing away any unbound substances, an enzyme-linked monoclonal antibody specific for ET-1 is added to the wells. Following a wash to remove any unbound antibody-enzyme reagent, a substrate solution is added to the wells and color develops in proportion to the amount of ET-1 bound in the initial step. The color development is stopped and the intensity of the color is measured.

SKU: USE3650-03 Categories: , ,

Additional information

Kit Components

E3650-03A: Microtiter Strips, 1×96 wells
E3650-03B: Endothelin-1 Mab (HRP), 1x21ml
E3650-03C: Standard, 1x1vial
E3650-03D: Assay Diluent, 1x18ml
E3650-03E: Calibrator Diluent, 1x21ml
E3650-03F: Wash Buffer, 25X, 1x21ml
E3650-03G: Color Reagent A, 1x12ml
E3650-03H: Color Reagent B, 1x12ml
E3650-03J: Stop Solution, 2N H2SO4, 1x6ml

Specificity

This assay recognizes natural and synthetic Endothelin-1.
The factors listed below were prepared at 250pg/ml in Calibrator Diluent and assayed for crossreactivity. Preparations of the following factors at 250pg/ml in a low level control were assayed for interference. No significant crossreactivity or interference was observed.

Human Peptides

Big Endothelin-1 (aa16-38), Big Endothelin-2, Endothelin-2, Sarafotoxin S6b, Sarafotoxin S6c

Rat Peptides

Big Endothelin-39

Bovine Peptides

Big Endothelin-39

Crossreactivity

Human Endothelin-2 shows 23.4% crossreactivity in this assay.

Human/Rat Endothelin-3 shows 0.5% crossreactivity in this assay.

Sensitivity

Thirty-four assays were evaluated and the minimum detectable dose (MDD) of Endothelin-1 ranged from 0.031-0.207pg/ml. The mean MDD was 0.087pg/ml.
The MDD was determined by adding two standard deviations to the mean optical density value of twenty zero standard replicates and calculating the corresponding concentration.

Precision

Intra-assay Precision:
Three samples of known concentration were tested 20 times on one plate to assess intra-assay precision.

Mean (pg/ml): 3.00, 7.34, 14.7
Standard Deviation: 0.120, 0.170, 0.280
CV (%): 4.0, 2.3, 1.9

Inter-assay Precision:
Three samples of known concentration were tested in 40 separate assays to assess inter-assay precision.

Mean (pg/ml): 3.05, 7.43, 14.4
Standard Deviation: 0.231, 0.438, 0.759
CV (%): 7.6, 5.9, 5.3

Storage and Stability

Store kit components at 4°C. Stable for 6 month after receipt at 4°C. After opening/reconstitution of reagents, stable for 1 month. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.

Shipping Temperature

On Dry Ice

Source Antigen

Human

Shelf Life

6 Months