< 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 Human sL-Selectin Immunoassay is a 2 hour solid phase ELISA that is designed to measure sL-Selectin in cell culture supernates, serum, and plasma. It contains recombinant human sL-Selectin and antibodies raised against the recombinant factor. This immunoassay has been shown to accurately quantitate the recombinant factor. Results obtained using natural human sL-Selectin showed linear curves that were parallel to the standard curves obtained using the recombinant kit standards. These results indicate that this kit can be used to determine relative mass values for natural human sL-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.
The recovery of sL-Selectin, spiked to three different levels ranging from 475-3784 ng/mL in various matrices, was evaluated.
Average % Recovery
Cell Culture Media (n=2)
EDTA Plasma (n=5)
To assess linearity of the assay two cell culture supernates and five spiked serum and plasma samples were diluted in Sample Diluent and assayed in three batches of kits.
Preparation and Storage
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, samples, and Control 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.
100 µL Standard, Control, or sample
Add 100 µL of Standard, Control, or sample to each well. Ensure sample addition is uninterrupted and completed within 15 minutes. Cover the plate with a plate sealer, and incubate at room temperature for 1 hour.
100 µL Conjugate
Add 100 µL of Conjugate to each well. Tap the plate gently to mix. Cover with a new plate sealer, and incubate at room temperature for 30 minutes.
Aspirate or decant each well and wash, repeating the process 5 times for a total of 6 washes.
100 µL Substrate
Add 100 µL of Substrate to each well. Cover with a new plate sealer, and incubate at room temperature for 30 minutes.
100 µL Stop Solution
Add 100 µL Stop Solution to each well. The Stop Solution should be added to the wells in the same order as the Substrate. Read at 450 nm within 30 minutes. Set wavelength correction to 620 nm or 650 nm.
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