Human/Mouse/Rat Activin A Quantikine ELISA Kit Summary

Product Summary

The Quantikine Human/Mouse/Rat Activin A Immunoassay is a 5.0 hour solid-phase ELISA designed to measure Activin A in human, mouse, or rat cell culture supernates, serum, plasma, and saliva. It contains CHO cell-expressed recombinant human/mouse/rat Activin A and has been shown to accurately quantitate the recombinant factor. Results obtained using natural human/mouse/rat Activin A 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 expressed Activin A.

Preparation and Storage

Background: Activin A

Activins, members of the TGF-beta superfamily, are disulfide-linked dimeric proteins originally purified from gonadal fluids as proteins that stimulated pituitary follicle stimulating hormone (FSH) release. Activin proteins have a wide range of biological activities, including mesoderm induction, neural cell differentiation, bone remodeling, hematopoiesis and roles in reproductive physiology. Activin isoforms and other members of the TGF-beta superfamily exert their biological effects by binding to heteromeric complexes of a type I and a type II serine-threonine kinase receptor, both of which are essential for signal transduction.

Activins are homodimers or heterodimers of the various beta subunit isoforms, while inhibins are heterodimers of a unique alpha subunit and one of the various beta subunits. Five beta subunits (mammalian beta A, beta B, beta C, beta E and Xenopus beta D) have been cloned to date. The activin/inhibin nomenclature reflects the subunit composition of the proteins: Activin A (beta A - beta A), Activin B (beta B - beta B), Activin AB (beta A - beta B), Inhibin A (alpha - beta A) and Inhibin B (alpha - beta B).

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.