Human FOLR1 Quantikine ELISA Kit Summary

Product Summary

The Quantikine Human FOLR1 Immunoassay is a 4.5 hour solid-phase ELISA designed to measure human FOLR1 in cell culture supernates, serum, plasma, saliva, urine, and human milk. It contains CHO cell-expressed recombinant human FOLR1 and has been shown to accurately quantitate the recombinant factor. Results obtained using natural human FOLR1 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 FOLR1.

Preparation and Storage

Background: FOLR1

Folate Receptor 1 (FOLR1), also known as Folate Receptor alpha and Folate Binding Protein (FBP), is a 37-42 kDa protein that mediates the cellular uptake of folic acid and reduced folates. Dietary folates are required for many key metabolic processes including nucleotide and methionine synthesis, the interconversion of glycine and serine, and histidine breakdown. Mature FOLR1 is an N-glycosylated protein that is anchored to the cell surface by a GPI linkage. Human FOLR1 shares 83 percent amino acid sequence identity with mouse and rat FOLR1. FOLR1 is predominantly expressed on epithelial cells and is dramatically up-regulated on many carcinomas. It is critically required during early embryogenesis as shown in knockout mice which die in utero with gross morphological defects. FOLR1 is internalized to the endosomal system where it dissociates from its ligand before recycling to the cell surface. A soluble form of FOLR1 can be proteolytically shed from the cell surface into the serum and breast milk.

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.