Human Progranulin Quantikine ELISA Kit Summary

Product Summary

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

Preparation and Storage

Background: Progranulin/PGRN

Progranulin, also known as Acrogranin, PCDGF, and Epithelin/Granulin Precursor, is a widely expressed glycoprotein that can undergo proteolysis to generate numerous Granulin peptides. Progranulin plays a role in the regulation of mesothelial differentiation, macrophage development, sexual differentiation of the brain, cartilage proliferation, wound repair, and the development of rheumatoid arthritis and osteoarthritis. It is often upregulated in cancer and contributes to tumor progression. Progranulin upregulation or mutation is associated with several neurological disorders including frontotemporal dementia, Creutzfeldt-Jakob disease, Alzheimer’s disease, and motor neuron disease.

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.