Human MIA Quantikine ELISA Kit

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DMIA00
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Human MIA ELISA Standard Curve
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Human MIA Quantikine ELISA Kit Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Assay Length
4.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (50 uL), Serum (25 uL), EDTA Plasma (25 uL), Heparin Plasma (25 uL), Saliva (50 uL)
Sensitivity
8.46 pg/mL
Assay Range
15.6 - 1,000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Saliva)
Specificity
Natural and recombinant human MIA
Cross-reactivity
< 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
Interference
No significant interference observed with available related molecules.

Product Summary

The Quantikine Human MIA Immunoassay is a 4.5 hour solid-phase ELISA designed to measure MIA in cell culture supernates, serum, plasma, and saliva. It contains E. coli-expressed recombinant Human MIA and antibodies raised against the recombinant factor. This immunoassay has been shown to accurately quantitate recombinant human MIA. Results obtained using natural human MIA showed linear curves that were parallel to the standard curves obtained using the Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for natural human MIA.

Precision

Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision
Inter-Assay Precision (Precision between assays) Three samples of a known concentration were tested in twenty separate assays to assess inter-assay precision. Assays were performed by at least three technicians using two lots of kit components

Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Saliva

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean 158 315 622 150 308 627
Standard Deviation 5.5 10.9 17.8 7.02 15.7 37.8
CV% 3.5 3.5 2.9 4.7 5.1 6

Recovery

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
Saliva (n=4) 102 86-110
Serum (n=4) 101 91-118

Linearity

To assess the linearity of the assay, samples containing and/or spiked with high concentrations of MIA were serially diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay. Samples were diluted prior to assay.
Human MIA ELISA Linearity

Data Examples

Human MIA ELISA Standard Curve

Product Datasheets

Preparation and Storage

Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: MIA

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.

Long Name:
Melanoma Inhibitory Activity
Entrez Gene IDs:
8190 (Human); 12587 (Mouse)
Alternate Names:
CD-RAP; Melanoma inhibitory activity protein; melanoma inhibitory activity; melanoma-derived growth regulatory protein; MIA
⚠ WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to www.P65Warnings.ca.gov.

Assay Procedure

Refer to the product for complete assay procedure.

Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
  1.   Prepare all reagents, standard dilutions, and samples as directed in the product insert.
  2.   Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.

  3. 50 µL Assay Diluent
  4.   Add 50 µL of Assay Diluent to each well.

  5. 50 µL Standard, Control, or Sample
  6.   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.
  7.   Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.

  8. 200 µL Conjugate
  9.   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.
  10.   Aspirate and wash 4 times.

  11. 200 µL Substrate Solution
  12.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.

  13. 50 µL Stop Solution
  14.   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.

FAQs

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