Human Neuropilin-1 Quantikine ELISA Kit Summary

Product Summary

The Quantikine Human Neuropilin-1 Immunoassay is a 4.5 hour solid-phase ELISA designed to measure human Neuropilin-1 in cell culture supernates, cell lysates, serum, plasma, urine, and human milk. It contains NS0-expressed recombinant human Neuropilin-1 and has been shown to accurately quantitate the recombinant factor. Results obtained using natural human Neuropilin-1 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 for naturally occurring human Neuropilin-1.

Preparation and Storage

Background: Neuropilin-1

Neuropilin-1 (Npn-1, previously known as neuropilin) and Neuropilin-2 (previously known as Npn-1-related molecule) are type I transmembrane proteins that bind distinct members of the class III secreted semaphorin subfamily that are implicated in repulsive axon guidance. Neuropilin extracellular domains contain two CUB (complement-binding) domains, two domains with homology to coagulation factors V and VIII, and a MAM (meprin) domain. In the absence of ligands, neuropilins can form homo- and hetero-oligomers via their MAM domains. Expression in developing neurons of the central and peripheral nervous systems is somewhat overlapping but distinct. Npn-1 and Npn-2 are also receptors for VEGF165 on endothelial and tumor cells.

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.