Human MIA Quantikine ELISA Kit Summary
Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Saliva
|Intra-Assay Precision||Inter-Assay Precision|
The recovery of MIA spiked to levels throughout the range of the assay in various matrices was evaluated.
|Sample Type||Average % Recovery||Range %|
|Cell Culture Media (n=4)||103||101-107|
|EDTA Plasma (n=4)||98||85-115|
|Heparin Plasma (n=4)||99||85-113|
Human MIA ELISA Standard Curve
Preparation and Storage
MIA, also known as cartilage-derived retinoic acid-sensitive protein (CD-RAP), is a secreted protein expressed in cartilaginous tissue during development and in the adult. MIA was identified based on the fact that it is highly expressed in malignant melanomas, but not in normal melanocytes. MIA has been shown to interact with fibronectin, and promotes invasion and metastasis of melanoma cells, perhaps by blocking integrin-fibronectin interactions.
Assay ProcedureRefer to the product
- Prepare all reagents, standard dilutions, and samples as directed in the product insert.
- Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.
- Add 50 µL of Assay Diluent to each well.
- Add 50 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours on a horizontal orbital microplate shaker.
- Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.
- Add 200 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours on the shaker.
- Aspirate and wash 4 times.
- Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.
- Add 50 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.
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