Human ULBP-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 ULBP-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: ULBP-1

ULBP-1, -2, and -3 were originally identified as ligands for the human cytomegalovirus glycoprotein UL16 and designated UL16-binding proteins (ULBP). These proteins are distantly related to major histocompatibility class I (MHC I) molecules, possessing the alpha 1 and alpha 2 Ig-like domains, but lacking the alpha 3 domain. Unlike MHC Class I, they have no capacity to bind peptide or interact with beta2-microglobulin. ULBP-1, -2, and -3 are known to bind to human NKG2D, an activating receptor expressed on NK cells, NKT cells, gamma delta T cells, and CD8+ alpha beta T cells.

MULT1 (mouse UL16-binding protein-like Transcript 1) is a 53 kDa, MHC Class I-like molecule that belongs to the mouse family of NKG2D ligands. It is a type I transmembrane glycoprotein that is synthesized as a 334 amino acid (aa) precursor. It contains a 25 aa signal sequence, a 186 aa extracellular region, a 19 aa transmembrane segment and a 104 aa cytoplasmic tail. The extracellular region contains an alpha-1 and alpha-2 like domain with two intrachain disulfide bonds.

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.