Human FABP4 Quantikine ELISA Kit Summary

Product Summary

The Quantikine Human FABP4 Immunoassay is a 4.5 hour solid-phase ELISA designed to measure human FABP4 in cell culture supernates, cell lysates, serum, and plasma. It contains E. coli-expressed recombinant human FABP4 and has been shown to accurately quantitate the recombinant factor. Results obtained using natural human FABP4 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 FABP4.

Preparation and Storage

Background: FABP4/A-FABP

FABP4 (Fatty Acid Binding Protein 4), also known as A-FABP and aP2, is the predominant FABP found in adipocytes and is often used as a marker for adipocyte differentiation. It is also expressed in macrophages, dendritic cells, and endothelial cells. FABP4 is a key mediator of intracellular fatty acid transport and metabolism in adipose tissue. Its expression is regulated by multiple factors including fatty acids, PPAR gamma agonists, and Insulin, and its levels increase with lipolytic stimulation. It can increase the hydrolytic activity of Hormone-Sensitive Lipase and increase the production of proinflammatory cytokines and chemokines. FABP4 upregulation in adipocytes and macrophages is associated with the development of Insulin resistance, hypertriacylglycerolaemia, and atherosclerosis. Serum levels of FABP4 are elevated in obesity and metabolic syndrome, type 2 diabetes, HIV-associated lipodystrophy, polycystic ovary syndrome, nonalcoholic fatty liver disease, atherosclerosis, and acute ischaemic stroke. FABP4 is also overexpressed in multiple cancer types including ovarian, bladder, glioblastoma, and oral, and it may play a role in tumor progression.

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.