| TiterZyme® Kits
(ELISA Kits) |
IMPORTANT: We have
recently (July 22, 2005) developed a new human Osteopontin kit (Catalog
No. 900-142) to replace our previous kit (Catalog No. 900-027).
Our new kit provides improved performance characteristics that
will provide superior results. The following Technical Bulletin
outlines the differences between the two kits:
Osteopontin Technical Bulletin
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|
Osteopontin Colorimetric (EIA) Kits |
| Assay Format |
Catalog # |
Kit Insert |
MSDS |
Assay Layout |
| human: 96 Well Kit |
900-142
|
|
|
|
| mouse: 96 Well Kit |
900-090 |
|
|
|
| rat: 96 Well Kit |
900-089 |
|
|
|
|
|
FOR RESEARCH PURPOSES ONLY, NOT FOR DIAGNOSTIC USE. |
 
|
Non-Radioactive
Assay Ranges:
human Osteopontin: 2-32 ng/mL
mouse Osteopontin: 1,000-64,000 pg/mL
rat Osteopontin: 74.0-4,750 pg/mL
Time to Answer:
human Osteopontin: 3 Hours
mouse Osteopontin: 2 Hours
rat Osteopontin: 2 Hours
|
Osteopontin (OPN) is an extracellular glycosylated bone phosphoprotein
with a polypeptide backbone of about 32,000 kDa that has been
independently discovered several times. It binds calcium and interacts
with the vitronectin receptor, avbeta3 integrin.
It has been shown to bind covalently to fibronectin via
transglutaminase-catalyzed cross-linking. In bone it is produced by
matrix-producing osteoblasts, at the mineralization front, and by bone
resorbing osteoclasts. In vivo, elevation in OPN levels are seen during
sepsis and metastatic cancers, and OPN production is induced by phorbol
esters. The production of OPN in cell culture is modulated by certain
cytokines, such as
IL-2, TGF-beta, EGF and PDGF. OPN is chemotactic for smooth muscle
cells and macrophages. OPN inhibits inducible nitric oxide synthase in
vascular tissue and cells transfected with OPN RNA have reduced
tumorigenicity.
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|
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1. Pipet Standards and Samples in duplicate into the wells and incubate.
2. Aspirate and wash the plate.
3. Pipet Antibody into the wells and incubate.
4. Aspirate and wash the plate.
5. Pipet Substrate into the wells and incubate.
6. Pipet Stop Solution and read on a plate reader at 450 nm.
7. Calculate sample concentrations from Standard Curve.
|
| |
human Osteopontin
(900-142) |
mouse Osteopontin
(900-090) |
rat Osteopontin
(900-089) |
| RANGE |
2-32 ng/mL
|
1,000-64,000 pg/mL
|
74.0-4,750 pg/mL
|
| SAMPLE SIZE |
100 µL |
100 µL |
100 µL |
| SAMPLES PER 96
WELL KIT |
41 in duplicate
|
39 in duplicate
|
39 in duplicate
|
| SENSITIVITY |
0.110 ng/mL
|
192.3 pg/mL
|
19.5 pg/mL
|
| PRECISION |
3.2-5.0% Intra
8.3-10.1% Inter |
4.1-5.0% Intra
8.7-9.5% Inter |
4.5-5.5% Intra
3.2-5.6% Inter |
SAMPLE TYPE
recommended dilution |
MDA-MB-435 Conditioned
Media ( 1:2)
urine (1:32)
EDTA plasma (1:8)
human milk (1:6400)
|
culture media (1:2)
EDTA plasma (1:100)
urine (1:3,000)
|
culture media (1:2)
EDTA plasma (1:10)
urine (1:1,000)
|
| TIME TO ANSWER |
3 Hours |
2 Hours |
2 Hours |
| SPECIES
SPECIFICITY |
human |
mouse |
rat |
|
| human |
| Compound |
Cross Reactivity |
Compound |
Cross Reactivity |
| human Osteopontin |
100% |
native human Osteopontin |
111% |
| native bovine Osteopontin |
< 0.1% |
recombinant mouse Osteopontin |
< 0.1% |
| |
| mouse |
| Compound |
Cross Reactivity |
Compound |
Cross Reactivity |
| mouse Osteopontin |
100% |
rat Osteopontin |
14.8% |
| human Osteopontin |
4.3% |
|
|
| |
| rat |
| Compound |
Cross Reactivity |
Compound |
Cross Reactivity |
| rat Osteopontin |
100% |
mouse Osteopontin |
<0.1% |
| human Osteopontin |
<0.1% |
|
|
+32 (0) 16
58 90 45 
+32 (0) 16
50 90 45