Using Griess Reagent System to measure Nitrite release from BV-2 cells

February 25, 2015

University of Nebraska Medical Center
Pharmacology and Experimental Neuroscience
Graduate Research Assistant

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Company:

Promega

Product Name:

Griess Reagent System

Catalog Number:

g2930

The goal of my research is to stimulate BV-2 cells (microglial cell line) with N-4YSyn (tail end of the antigenic protein found in Parkinson's disease) and then evaluate the BV-2 cells for increased nitrite production.

Experimental Design and Results Summary

Application

Absorbance measured by plate reader

Starting Material

Cell line

Protocol Overview

BV-2 cells are co-cultured with varying concentrations of antigen (N-4YSyn) for 18-48 hours. Stimulating media is then removed from cells and replaced with regular cell media. Nitrite is then measured using Griess Reagent System from 0-12 hours post media change.

Tips

Be careful if your stimulating antigen contains nitrogen as this can effect the results of the assay.

Results Summary

The antigen N-4YSyn has the ability to increase the release of nitrite form BV-2 cells in a concentration-dependent manner, with peak nitrite release seen at 100 micrograms/ml of antigen.

Additional Notes

None

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Summary

The Good

It is very easy to use

The Bad

It does not seem to be able to detect the presence of very small amounts of nitrite.

The Bottom Line

This kit is very inexpensive, can be used for up to ten 96-well plates, and has a fairly long shelf life so if you run at least 2 plates a month using this assay it will not expire on you... it's a great bang for your buck!

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