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Mouse Endocan Quantikine ELISA Kit

Catalog #: MEND00 Datasheet / COA / SDS

Discontinued Product

MEND00 has been discontinued.
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Summary Precision Recovery Linearity Product Datasheets Preparation and Storage Background Related Research Areas

Mouse Endocan Quantikine ELISA Kit Summary

Assay Length
4.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (50 uL), Serum (20 uL), EDTA Plasma (20 uL), Heparin Plasma (20 uL)
Sensitivity
21.3 pg/mL
Assay Range
46.9 - 3,000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma)
Specificity
Natural and recombinant mouse Endocan
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 Mouse Endocan immunoassay is a 4.5 hour solid-phase ELISA designed to measure Endocan in cell culture supernates, mouse serum, and plasma. It contains NS0-expressed recombinant mouse Endocan and antibodies raised against the recombinant factor. This immunoassay has been shown to accurately quantitate the recombinant mouse Endocan. Results obtained using natural mouse Endocan showed dose response curves that were parallel to the standard curves obtained using the Quantikine mouse kit standards. These results indicate that this kit can be used to determine relative mass values for natural mouse Endocan.

Precision

Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in separate assays to assess inter-assay precision

Cell Culture Supernates

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 40 40 40
Mean (pg/mL) 143 398 1107 137 373 1097
Standard Deviation 14.1 25.3 70.7 13.1 23.2 78.4
CV% 9.8 6.4 6.4 9.6 6.2 7.1

Serum, EDTA Plasma, Heparin Plasma

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 40 40 40
Mean (pg/mL) 154 515 1439 206 517 1406
Standard Deviation 14.8 25.4 86 20.8 39.2 99.8
CV% 9.6 4.9 6 10.1 7.6 7.1

Recovery

The recovery of mouse Endocan spiked to three levels throughout the range of the assay was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Samples (n=4) 99 91-120
EDTA Plasma (n=4) 92 81-119
Heparin Plasma (n=4) 108 89-119
Serum (n=4) 92 81-117

Linearity

To assess the linearity of the assay, samples containing or spiked with high concentrations of mouse Endocan in each matrix were diluted with the appropriate Calibrator Diluent and assayed.

Product Datasheets

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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 -20 to -70 °C. Use a manual defrost freezer and avoid repeated freeze-thaw cycles. Do not use past expiration date.

Background: Endocan/ESM-1

Endocan, also known as Endothelial Cell-specific Molecule-1 (ESM-1) and IGFBP-rp6, is a secreted cysteine-rich dermatan sulfate proteoglycan. It is primarily expressed by endothelial cells within the vascular network of the kidney and in the alveolar walls of the lung. Endocan expression is upregulated by TNF-alpha, IL-1 beta and LPS, and downregulated by IFN-gamma. Endocan binds LFA-1 on human lymphocytes, monocytes and Jurkat cells, inhibiting its binding to ICAM-1 and reducing LFA-1-mediated leukocyte activation. Elevated circulating Endocan levels have been reported in patients with lung cancer and severe sepsis.

Entrez Gene IDs:
11082 (Human); 71690 (Mouse)
Alternate Names:
Endocan; endothelial cell-specific molecule 1; ESM1; ESM-1; IGFBP-rp6

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
  3.   Add 50 µL of Assay Diluent to each well.
    4. 50 µL Standard, Control, or Sample
  4.   Add 50 µL of Standard, Control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.
  5.   Aspirate each well and wash, repeating the process 4 times for a total of 5 washes.
    6. 100 µL Conjugate
  6.   Add 100 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.
  7.   Aspirate and wash 5 times.
    8. 100 µL Substrate Solution
  8.   Add 100 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.
    9. 100 µL Stop Solution
  9.   Add 100 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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