Human GM-CSF Quantikine ELISA Kit Summary

Product Summary

The Quantikine Human GM-CSF Immunoassay is a 3.5 or 4.5 hour solid phase ELISA designed to measure GM-CSF levels in cell culture supernates, serum, and plasma. It contains E. coli-expressed recombinant human GM-CSF and antibodies raised against the recombinant factor. It has been shown to quantitate recombinant human GM-CSF accurately. Results obtained using natural human GM-CSF 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 natural GM-CSF.

Preparation and Storage

Background: GM-CSF

GM-CSF (Granulocyte-Macrophage Colony Stimulating Factor) induces the development of monocytes, neutrophils, eosinophils, and myeloid and dermal dendritic cells. It also acts as a neutrophil and dendritic cell chemoattractant. GM-CSF promotes Th1 and Th17 cell mediated autoimmune inflammation as well as the inflammatory activation of dendritic cells, microglia, alveolar macrophages, and eosinophils. In addition, it cooperates with G-CSF in promoting tumor cell proliferation and invasion. GM-CSF signals through a receptor complex composed of GM-CSF R alpha and the Common beta Chain (beta c) with Syndecan-2 as a potential co-receptor. The beta c subunit is shared by the receptor complexes for IL-3 and IL-5.

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.