Human Glypican 3 Quantikine ELISA Kit

Product Summary

The Quantikine Human Glypican 3 Immunoassay is a 4.5 hour solid phase ELISA designed to measure Glypican 3 levels in cell culture supernates, cell lysates, serum, and plasma. It contains NS0-expressed recombinant human Glypican 3 and antibodies raised against the recombinant protein. Results obtained for naturally occurring human Glypican 3 showed linear curves that were parallel to the standard curves obtained using the Quantikine Human Glypican 3 Immunoassay standards. These results indicate that this kit can be used to determine relative mass values for natural human Glypican 3.

Preparation and Storage

Background: Glypican 3

The glypicans (GPC) constitute a family of glycosylphosphatidylinositol (GPI)-anchored, heparan sulfate proteoglycans. Six members of this family have been identified in mammals (GPC1-GPC6). All glypican core proteins contain an N-terminal signal peptide, a large globular cysteine-rich domain (CRD) with 14 invariant cysteine residues, a stalk-like region containing the heparan sulfate attachment sites, and a C-terminal GPI attachment site.

Mutations in Glypican 3 cause a rare disorder in humans, Simpson-Golabi-Behmel Syndrome, which is characterized by pre- and postnatal overgrowth of multiple tissues and organs and an increased risk for developing embryonic tumors. These features are also present in the mouse knock-out of Glypican 3 indicating that Glypican 3 regulates cell survival and inhibits cell proliferation during development. Glypican 3 has been implicated in regulating many different signaling pathways including: IGF, FGF, BMP and Wnt. An endoproteolytic processing of Glypican 3 by proprotein convertases is required for the modulation of Wnt signaling. Direct interaction of Glypican 3 with FGF-basic has been observed and is mediated by the heparan sulfate chains.

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.