< 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
No significant interference observed with available related molecules.
The Quantikine Mouse soluble E-Selectin (sE-Selectin) Immunoassay is a 4.5 hour solid phase ELISA designed to measure mouse sE-selectin levels in cell culture supernates, serum, and EDTA plasma. It contains NS0-expressed recombinant mouse sE-selectin and antibodies raised against the recombinant protein. Results obtained for naturally occurring mouse sE-selectin 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 of natural mouse sE-selectin.
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 known concentration were tested in separate assays to assess inter-assay precision
Cell Culture Supernates, Serum, EDTA Plasma
The recovery of mouse sE-Selectin spiked to three levels throughout the range of the assay in various matrices was evaluated.
Average % Recovery
Cell Culture Supernates (n=6)
EDTA Plasma (n=4)
To assess the linearity of the assay, five or more samples containing and/or spiked with high concentrations of mouse sE-Selectin in each matrix were diluted with Calibrator Diluent and then assayed.
Store the unopened product at 2 - 8 °C. Do not use past expiration date.
The Selectin family is comprised of three members, E-Selectin, L-Selectin, and P-Selectin. E-Selectin [endothelial leukocyte adhesion molecule-1 (ELAM-1), CD62E] is transiently expressed on vascular endothelial cells in response to IL-1 beta and TNF-alpha. The human and rat proteins share approximately 67% amino acid sequence identity. The mouse and rat proteins share approximately 78% amino acid sequence identity. L-Selectin (Leukocyte Selectin, LAM-1, LECAM-1, LECCAM-1, TQ1, Leu-8, MEL-14 antigen, DREG, lymph node homing receptor, CD62L) is expressed constitutively on a wide variety of leukocytes. Two forms of L-Selectin have been reported, apparently arising as a result of post-translational modifications. Human and mouse L-Selectin share 76% amino acid sequence identity. Human P-Selectin (GMP-140, LECAM-3, PADGEM, CD62P) is expressed by activated platelets and endothelial cells. The extracellular domains of human and mouse P-Selectin share approximately 73% amino acid sequence identity.
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.
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.
50 µL Assay Diluent
Add 50 µL of Assay Diluent to each well.
50 µL Standard, Control, or Sample
Add 50 µL of Standard, Control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.
Aspirate each well and wash, repeating the process 4 times for a total of 5 washes.
100 µL Conjugate
Add 100 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.
Aspirate and wash 5 times.
100 µL Substrate Solution
Add 100 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.
100 µL Stop Solution
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.
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