|Method:||Sandwich ELISA, HRP/TMB, 12x8-well strips|
|Sample type:||Urine, serum, plasma (EDTA, heparin, citrate)|
|Standard range:||0-800 pmol/l (7 standards)|
|Standard points:||0/25/50/100/200/400/800 pmol/l|
|Sample size:||10-20 µl / well|
|Incubation time:||3 h / 1 h / 30 min|
|Unit conversion:||1 pg/ml = 0.05 pmol/l (MW: 20 kDa)|
LOD: 4 pmol/l (0 pmol/l + 3 SD), LLOQ: 3 pmol/l
Intra-assay (n=5) ≤ 6%, Inter-assay (n=16) ≤ 5%
The mean recovery of recombinant Endostatin in serum and citrate plasma is 95% and 91%, respectively.
The mean recovery of recombinant Endostatin in heparin plasma and EDTA plasma is 96% and 92%, respectively.
The mean dilution linearity of endogenous Endostatin in serum sampes is:
Mean for 1+29 dilution = 119%
Mean for 1+59 dilution = 120%
Mean for 1+119 dilution = 116%.
This assay recognizes endogenous and recombinant human Endostatin. There is no cross reactivitiy with other species. There is no cross reactivity with human recombinant Collagen Type XV (COL15).
Values from apparently healthy individuals:
Median (serum, n=59): 5.1 nmol/l.
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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Endostatin, a 20-kDa C-terminal proteolytic fragment of collagen XVIII, is an endogenous angiogenesis inhibitor localized in the vascular basement membrane in various organs (http://www.uniprot.org/uniprot/P39060). The biological functions of the endostatin-network involve SPARC, thrombospondin-1, glycosaminoglycans, collagens, and integrins.
Endostatin is expressed during the progression of renal fibrosis in tubular cells of injured tissue. In renal microvascular disease, observed in late stages of patients with chronic kidney disease, increased endostatin levels are possibly the consequence of enhanced extracellular matrix degradation. Thus endostatin may become an important marker for progressive microvascular renal disease in patients with chronic kidney disease. Endostatin levels in blood are also likely to increase in patients with other microvascular tissue injuries, including atherosclerosis, myocardial and brain ischemia. In ischemic stroke patients, high endostatin plasma levels predict a worse long-term clinical outcome.
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