Capture ELISA Kit To Measure Human BFGF (Basic FGF)

The Scripps Research Institute
The Cell and Molecular Biology
Assistant Professor

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Company:

PeproTech

Product Name:

Human FGF-basic Standard ABTS ELISA Development Kit

Catalog Number:

900-K08

Basic FGF (bFGF) is one of the major growth factors that stimulate the formation of new (angiogenic) blood vessels. Measurement of bFG production by tumor cells is important in the research focused on tumor angiogenesis and cancer metastasis. Since growth factor production by mammalian cells is relatively low (usually from picograms to nanograms per ml of conditioned medium or tumor extract),  a sandwich (capture) ELISA format is required for the quantitative measurement of naturally expressed cytokines.  We employed the PeproTech bFGF ELISA kit to measure and compare the levels of bFGF secreted by human tumor cells exhibiting different levels of metastatic dissemination in vivo.

Experimental Design and Results Summary

Application

Sandwich (capture) ELISA

Starting Material

bFGF was measured in the extracts from primary human tumors treated with aprotinin or vehcle (PBS). The kit contains all key components to quantify the levels of bFGF by a standard caprure ELISA.

Protocol Overview

We closely followed the manufacturer's "ELISA: Sandwich ABTS Protocol", but with some modifications indicated in the "tips" section.

Tips

The anti bFGF "capture" antibody could be coated at 0.5 microgram per ml, instead of recommended 1 microgram per ml. Coating of the "capture" antibody could be done at 4oC instead of RT. Washings could be done 3 times instead of recommended 4 times.

Results Summary

Yellow color substrate precipitates in the wells containing captured bFGF. Color development is measured with a spectrophotometer (ELISA plate reader) at 405 nm. The optical density is proportional to the levels of bFGF.

DOI or PMID #

N/A

Additional Notes

The image depicts the standard bFGF curve (the graph on the left) that is used to quantify bFGF concentrations in tumor samples (the 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 human bFGF kit from PeproTech is an excellent sandwich (capture) ELISA kit to quantitatively measure the levels of bFGF produced by human tumor cells.

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