Cholesterol Fluorometric Assay Kit

March 25, 2015

Hershey Medical Center PSU
Neurosurgery
Research tech

Overall

Quality of Results

Ease-of-Optimization

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Company:

BioVision

Product Name:

Cholesterol Fluorometric Assay Kit

Catalog Number:

K603-100

The general research aim for using this cholesterol kit was to see if certain cell lines had cholesterol and then to compare between cell lines. I also used it to measure cholesterol once I treated the cells with certain chemicals. This kit seemed easy to use and was a fairly quick assay. Also the kit was fluorometric. I was able to get very good results from this kit and have recommended this kit to other lab members and they had similar feelings with using this kit.

Experimental Design and Results Summary

Application

Detecting cholesterol in cell lines

Starting Material

Cell lysates

Protocol Overview

1. Plated cells to certain density 2. Prepped the cells for lysates according to kit protocol 3. Added the mixture of: cholorform, isopropanol, NP-40 4. Sonicated 5. Centrifuged 6. Transferred liquid to new tube 7. Speed vacced the samples 8. Resuspended the pellet with cholesterol assay buffer

Tips

Be sure when first starting to do the dilution factor the manufacteur suggests

Results Summary

The results were in OD readings and then you had to use the formulas that they gave you in the protocol and then plot them and then convert them to ug/ug of protein.

Additional Notes

None

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Summary

The Good

Fairly quick and easy to use

The Bad

Various calculations needed

The Bottom Line

This kit was easy to use. There were many calculations, but once we got one done, the others were easier and didn't take as long since this was used as a template.

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