|Method:||Sandwich ELISA, HRP/TMB, 12x8-well strips|
|Sample type:||Serum, EDTA plasma|
|Standard range:||0-10 fmol/ml (6 serum based standards)|
|Standard points:||0/0.625/1.25/2.5/5/10 fmol/ml|
|Controls:||2 serum based controls|
|Sample size:||50 µl / well|
|Incubation time:||overnight / 1 h / 30 min|
|Unit conversion:||1 pg/ml = 0.4 fmol/ml (MW: 2.49 kDa)|
LOD: 0.02 fmol/ml (0 fmol/ml + 3 SD)
Intra-assay (n=18) ≤ 4%, Inter-assay (n=24) ≤ 6%
The mean recovery of recombinant Endothelin (spike = 1 fmol/ml) is 96%.
The mean recovery of recombinant Endothelin (spike = 5 fmol/ml) is 86%.
ET(1-21): 100%, ET2 (1-21): 100%, ET3 (1-21): <5%, Big Endothelin (1-38): <1%, Big Endothelin ( 22-38): <1%.
In normal human plasma samples ET-2 is estimated to be present at less than 20% of the ET-1 level. ET-3 is estimated to be present at 50% of the ET-1 level.
Animal sera: horse, cat, pig 100%; dog 66% and rat 79%, Mouse sera can not be measured in this ELISA.
Values from apparently healthy individuals:
Median (n=70): 0.26 fmol/ml.
It is recommended to establish the normal range for each laboratory.
Principle of the assay:
Manual ELISAs - easily adaptable for automation!
INSTRUCTIONS FOR USE
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Cleavage of Big Endothelin by a membrane-bound metalloproteinase, the Endothelin Converting Enzyme (ECE) leads to the active ET (1-21), a potent vasoconstrictor, and to the biological inactive C-terminal fragment (22-38). The half-life of ET in the plasma is less than one minute, whereas clearance of Big ET is much slower. Endothelin has been identified in a variety of tissues, including lung, kidney, brain, pituitary and peripheral endocrine tissues, and placenta. The biological role of ET extends beyond regulating vascular tone also in its growth regulatory properties. The peptide interacts in an autocrine/paracrine manner with specific ET receptors found on numerous cells, including smooth muscle cells, myocytes, and fibroblasts.
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