Human LYVE-1 DuoSet ELISA Summary

Product Summary

This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant human LYVE-1. The suggested diluent is suitable for the analysis of most cell culture supernate samples. Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet.

Preparation and Storage

Background: LYVE-1

Lymphatic Vessel Endothelial Hyaluronan (HA) Receptor-1 (LYVE-1) is a 60-kDa type I transmembrane glycoprotein that is a member of the Link Protein superfamily. HA is found in the extracellular matrix of most animal tissues and in body fluids. It modulates cell behavior and functions during tissue remodeling, development, homeostasis, and disease. It is often used as a marker of lymphatic endothelia.

LYVE-1 is expressed on both the lumenal and ablumenal surfaces of lymphatic endothelium, and also on hepatic blood sinusoidal endothelia. This expression pattern, combined with studies showing that LYVE-1 can support cellular HA internalization in vitro, may suggest LYVE-1 participation in HA internalization for degradation, or transport of HA from tissues into the lumen of lymphatic vessels. LYVE-1-directed HA localization to lymphatic surfaces might also affect aspects of the immune response or tumor metastases. HA binding to CD44 can still occur in the presence of LYVE-1 in vitro. Therefore, LYVE-1-directed HA localization to lymphatics could provide a substrate for transmigrating CD44+ leukocytes or tumor cells. In addition to hepatic and lymphatic endothelia, some expression of LYVE-1 has been reported on Kupffer cells, the islets of Langerhans, cortical neurons, and renal epithelium. Roles for LYVE-1 on these cell and tissue types remain to be determined.

Assay Procedure

Refer to the product datasheet for the 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、Add 50 μL of Assay Diluent to each well.

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、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、Add 100 μL Substrate Solution to each well.

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.