Human FABP2/I-FABP Quantikine ELISA Kit Summary

Product Summary

The Quantikine Human FABP2/I-FABP Immunoassay is a 4.5 hour solid phase ELISA designed to measure FABP2 levels in cell culture supernates, cell lysates, serum, plasma, and urine. It contains E. coli-expressed recombinant human FABP2 and antibodies raised against the recombinant protein. Results obtained for naturally occurring human FABP2 showed linear curves that were parallel to the standard curves obtained using the Quantikine Human FABP2 Immunoassay standards. These results indicate that this kit can be used to determine relative mass values for natural human FABP2.

Preparation and Storage

Background: FABP2/I-FABP

Fatty acid binding proteins are small cytoplasmic lipid binding proteins that are expressed in a tissue specific manner. FABPs bind free fatty acids, cholesterol, and retinoids, and are involved in intracellular lipid transport. Circulating FABP levels are used as indicators of tissue damage. Some FABP polymorphisms have been associated with disorders of lipid metabolism and the development of atherosclerosis.

FABP2, also known as intestinal fatty acid binding protein (I-FABP or FABPI) and gut FABP (gFABP), is a member of the cytosolic fatty acid binding protein family. FABP2 mediates the absorption and intracellular transport of dietary long-chain fatty acids. Genetic variations of FABP2 are implicated in obesity and Type II diabetes. Human FABP2 shares 78%, 82%, and 86% amino acid sequence identity with mouse, rat, and canine FABP2, respectively.

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.