Abstract
An ELISA was set up using polyvinylchloride microtiter plates coated with rabbit anti-UK IgG's and affino-purified goat anti-UK IgG's as second antibody. Detection occurred with rabbit anti-goat IgG antibodies conjugated with alkaline phosphatase. The assay is specific for urokinase (UK) with a detection limit of 100 pg/ml sample. Tissue-type plasminogen activator, up to concentrations of 100 ng/ml, does not interfere. The assay measures the antigen of the inactive zymogen pro-UK, the active enzyme UK and the UK-inhibitor complex with equal efficiency and gives the total UK antigen present, irrespective of its molecular form. Culture media of fibroblasts, endothelial- and kidney cells showed, despite the absence of active UK, antigen levels of 1.2, 23 and 65 ng/ml, respectively. In human plasma the UK concentration was found to be 3.5 +/- 1.4 ng/ml (mean +/- SD, n = 54). The inter- and intra-assay variations were 20% and 6%, respectively.
| Original language | English |
|---|---|
| Pages (from-to) | 569-577 |
| Journal | Thrombosis and haemostasis |
| Volume | 43 |
| Issue number | 5 |
| DOIs | |
| Publication status | Published - 1 Sep 1986 |
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Keywords
- Animals
- Antibodies
- Antigen-Antibody Complex
- Aorta
- Carcinoma, Squamous Cell
- Cell Line
- Cercopithecus aethiops
- Culture Media
- Endothelium
- Enzyme-Linked Immunosorbent Assay
- Fibroblasts
- Humans
- Isoflurophate
- Kidney
- Liver
- Melanoma
- Reference Values
- Urokinase-Type Plasminogen Activator
- Journal Article
- Research Support, Non-U.S. Gov't
Cite this
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Quantitation of urokinase antigen in plasma and culture media by use of an ELISA. / Binnema, D J; van Iersel, J J; Dooijewaard, G.
In: Thrombosis and haemostasis, Vol. 43, No. 5, 01.09.1986, p. 569-577.Research output: Contribution to journal › Article › Academic › peer-review
TY - JOUR
T1 - Quantitation of urokinase antigen in plasma and culture media by use of an ELISA
AU - Binnema, D J
AU - van Iersel, J J
AU - Dooijewaard, G
PY - 1986/9/1
Y1 - 1986/9/1
N2 - An ELISA was set up using polyvinylchloride microtiter plates coated with rabbit anti-UK IgG's and affino-purified goat anti-UK IgG's as second antibody. Detection occurred with rabbit anti-goat IgG antibodies conjugated with alkaline phosphatase. The assay is specific for urokinase (UK) with a detection limit of 100 pg/ml sample. Tissue-type plasminogen activator, up to concentrations of 100 ng/ml, does not interfere. The assay measures the antigen of the inactive zymogen pro-UK, the active enzyme UK and the UK-inhibitor complex with equal efficiency and gives the total UK antigen present, irrespective of its molecular form. Culture media of fibroblasts, endothelial- and kidney cells showed, despite the absence of active UK, antigen levels of 1.2, 23 and 65 ng/ml, respectively. In human plasma the UK concentration was found to be 3.5 +/- 1.4 ng/ml (mean +/- SD, n = 54). The inter- and intra-assay variations were 20% and 6%, respectively.
AB - An ELISA was set up using polyvinylchloride microtiter plates coated with rabbit anti-UK IgG's and affino-purified goat anti-UK IgG's as second antibody. Detection occurred with rabbit anti-goat IgG antibodies conjugated with alkaline phosphatase. The assay is specific for urokinase (UK) with a detection limit of 100 pg/ml sample. Tissue-type plasminogen activator, up to concentrations of 100 ng/ml, does not interfere. The assay measures the antigen of the inactive zymogen pro-UK, the active enzyme UK and the UK-inhibitor complex with equal efficiency and gives the total UK antigen present, irrespective of its molecular form. Culture media of fibroblasts, endothelial- and kidney cells showed, despite the absence of active UK, antigen levels of 1.2, 23 and 65 ng/ml, respectively. In human plasma the UK concentration was found to be 3.5 +/- 1.4 ng/ml (mean +/- SD, n = 54). The inter- and intra-assay variations were 20% and 6%, respectively.
KW - Animals
KW - Antibodies
KW - Antigen-Antibody Complex
KW - Aorta
KW - Carcinoma, Squamous Cell
KW - Cell Line
KW - Cercopithecus aethiops
KW - Culture Media
KW - Endothelium
KW - Enzyme-Linked Immunosorbent Assay
KW - Fibroblasts
KW - Humans
KW - Isoflurophate
KW - Kidney
KW - Liver
KW - Melanoma
KW - Reference Values
KW - Urokinase-Type Plasminogen Activator
KW - Journal Article
KW - Research Support, Non-U.S. Gov't
U2 - https://doi.org/10.1016/0049-3848(86)90077-0
DO - https://doi.org/10.1016/0049-3848(86)90077-0
M3 - Article
C2 - 3750278
VL - 43
SP - 569
EP - 577
JO - Thrombosis and haemostasis
JF - Thrombosis and haemostasis
SN - 0340-6245
IS - 5
ER -