Human FGF acidic/FGF1 Quantikine ELISA Kit Summary

Product Summary

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

Preparation and Storage

Background: FGF acidic/FGF1

FGF acidic, also known as FGF-1, ECGF, and HBGF-1, is a secreted mitogen that stimulates the proliferation of all cells of mesodermal origin and many cells of neuroectodermal, ectodermal, and endodermal origin. It plays a number of roles in development, regeneration, and angiogenesis. FGF acidic is released extracellularly as a disulfide-linked homodimer and is stored in complex with extracellular heparan sulfate. The association of FGF acidic with heparan sulfate is a prerequisite for its subsequent interaction with FGF receptors. Internalized FGF acidic can translocate to the cytosol and to the nucleus. Intracellular FGF acidic functions as a survival factor by inhibiting p53 activity and proapoptotic signaling.

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.