Capture ELISA Kit To Measure Human GM-CSF

The Scripps Research Institute
The Cell and Molecular Biology
Assistant Professor

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Company:

PeproTech

Product Name:

Human GN-CSF Mini ABTS ELISA Development Kit

Catalog Number:

900-M30

Granulocyte-macrophage colony stimulating factor (GM-CSF) is a cytokine inducing differentiation of hematopoietic stem cells into granulocytes and monocytes/macrophages. GM-CSF is also a critical regulator of inflammation and immune responses. In tumor microenvironment, GM-CSF acts as a potent chemoattractant for myeloid cells, especially neutrophils. We measured GM-CSF production in human tumor cell variants to correlate the extent of neutrophil infiltration of developing tumor xenografts with the metastatic potential of tumor cells. Since naturally-produced cytokines are secreted in very low amounts, their measurement in cell conditioned medium, tumor extracts or serum of tumor-bearing mice requires the use of a sandwich (capture) ELISA kits.

Experimental Design and Results Summary

Application

Sandwich (capture) ELISA

Starting Material

Samples of serum free medium conditioned by human tumor cells for 24 hr. The kit contains all key components to quantify the levels of GM-CSF by a standard caprure ELISA.

Protocol Overview

We closely follow the manufacturer's protocol for ABTS ELISAs with some small modifications. We coat capture antibody at 0.5 microgram/mL, overnight, 4oC.

Tips

Plates could be washed 3 times instead of recommended 4.

Results Summary

The color reaction develops in wells containing GM-CSF. Negative control wells remain almost colorless. The intensity of color is proportional to the amount of captured GM-CSF. The concentration of GM-CSF in the samples is calculated from the standard curve.

DOI or PMID #

N/A

Additional Notes

The image depicts the standard GM-CSF curve (the graph on the left) that was used to quantify GM-CSF secreted by tumor cells characterized low and high metastatic potentlial (graph on the right).

Image Gallery

Summary

The Good

The kit is very specific, sensitive, and easily adaptable.

The Bad

None detected.

The Bottom Line

The GM-CSF capture (sandwich) ELISA kit "900-M30" from PeproTech is an excellent tool to determine concentrations of naturally produced GM-CSF.

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