Crisp Clean Catalase

Overall

Quality of Results

Ease-of-Optimization

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Cardiovascular Innovation Institute
University of Louisville
Senior Research Technologist

Company:

Calbiochem

Product Name:

Anti-Catalase Rabbit pAb

Catalog Number:

219010

The stromal vascular fraction (SVF) from epididymal fat pads of rats have shown to possess vasculogenic and angiogenic potential in our research. However, upon comparing young rat SVF cell populations to old rat SVF cell populations there is a decreased vasculogenic and angiogenic potential seen in the aged SVF cell population compared to the young. In an attempt to determine potential contributing factors to these findings our lab wanted to investigate the protein levels of catalase in these SVF cell populations. Catalase is involved in preventing damage by reactive oxygen species related to hydrogen peroxide levels in cells so it was hypothesized that perhaps the old SVF samples had considerably less catalase and hence suffered greater hindrance from reactive oxygen species. This antibody was chosen because of its multiple references in research papers and the PI's previous success with this antibody in another lab before becoming a PI.

Experimental Design and Results Summary

Applications

Western Blot

Sample

Female Rat SVF Cell Samples

Primary Incubation

1:1000 in 1% Milk/TBS-T for 2 hours at room temperature

Blocking Agent

5% Milk protein in Tris-Buffered Solution + Tween (TBS-T) for 1 hour at room temperature on rocker

Secondary Incubation

Goat anti-rabbit IgG dylight 488 (cat# 35552) at 1ug/mL (1:1000) in 1% Milk/TBS-T for 1 hour at room temperature

Tertiary Incubation

N/A

Detection

Typhoon Blue Laser 488 526 filter 400 PMT

Results Summary

Catalase bands are crisp and clean without any bleed through between samples making for easy and reliable measurement of the densitometry. 25ug of protein were used for each sample to attain these results. This type of western was repeated several times and gave consistent results each time with the protocol outlined in this review.

Additional Notes

None

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Summary

The Good

Clean results, consistent results, cost effective

The Bad

The company's billing process. Otherwise none for the antibody itself.

The Bottom Line

Great antibody for western use. Our lab has not tried this antibody for any other applications at this time.

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