PLEASE help! Antioxidant assay kit.

[theroses]

Hello,
My science fair project is determining whether lifestyle changes affect telomere growth in humans. I will determine this by measuring the cortisol and antioxidant level in cells before and after a period of time. The problem is that I already have a possible kit for the cortisol assay, but none for the antioxidant assay.

Does anyone know of any good (relatively affordable) kits for antioxidant assay, or any other ways to measure antioxidant levels in human cells?

thank you.

[Adean2001]

Hello. I am afraid I haven't measured levels of antioxidants myself. A google of "antioxidant assay" brought up a number of results for kits you can buy (that are quite pricey...I think the cheapest one I saw was for over $200). I also found this article http://pac.iupac.org/publications/pac/p ... 5x0957.pdf that provides a number of antioxidant assays. Perhaps you can go through this article and google the various methods they discuss to see how involved the protocols are.

I think you will probably get better advice by posting this on the chemistry boards as this is really a chemistry question (even though overall topic is geared more toward life science). Sorry I couldn't be of more help.

[SciB]

Hi Roses,

I'm glad you are proceeding with the telomere project but I'm not sure I understand how cortisol and antioxidant levels will give you information about the effects of lifestyle on telomere length. Can you please explain that to me. You said you are using human cells for the experiments. What kind of human cells are you using and how are you treating them to create the effect of different life styles? How are you measuring cortisol, by ELISA?

The term 'antioxidant' is a broad and general category of molecules that specifically inhibit the production or promote the breakdown of reactive oxygen species (ROS). The three main cellular antioxidants are glutathione, peroxidase and superoxide dismutase.

Carolina Biologicals has a kit for detecting 'oxidases' but I'm not sure if that will work for you. I would call them and explain what you are doing and ask them if their kit is appropriate. Here's the URL: http://www.carolina.com/biological-medi ... eroxidasee

I checked some of the biotech companies for glutathione or antioxidant assay kits and they are all in the $375-400 range.

You have a couple of options--call companies and ask them to donate the reagents to you as a public service benefit for young students (some companies are generous this way) or ask around among labs at a university where they may have an old glutathione kit you could have. Most labs have expired ELISA kits that are still OK because they come with all the controls to verify activity.

Good luck and please keep ALL your posts in the original telomerase thread because we look for them under that title. I just found this new post by accident when I recognized your name.

Sybee

[theroses]

Thank you SciB, I went to the site you gave me and I do think that it is what I need.

Do you also know of any cortisol level in human cells kits? I'm not sure if the one I found will be affordable (the company I contacted did not respond yet).

Thank you.

[deleted-132180]

Hi theroses,

I really like the topic of how lifestyle can affect telomere length, but I'm curious as to how cortisol and/or antioxidant levels can have an effect on telomere length? Has there been any research done in this area that shows this? Furthermore, what types of cells are you going to measure these levels in? Are they going to be primary cells derived from actual animals or patients? Or are they going to be cell lines? How often are you going to measure the cortisol and/or antioxidant levels? And in addition to that, are you going to be measuring the average telomere lengths of your cells simultaneously to see if you can find a correlation between cortisol and/or antioxidant levels with telomere length? If so, how are you going to measure telomere length?

Another thing I'm afraid of is that if you're just going to measure cortisol and/or antioxidant levels from cultured cells over a period of time, I don't think there would be huge fluxes unless the cell is put under some sort of stress that can increase or decrease these levels. One thing you can probably try is to subject these cells to certain stresses and then see how that affects the length of their telomeres over time. Or you can somehow alter the cortisol and/or antioxidant levels in your cultured cells and see how increasing or decreasing the levels of these compounds can affect telomere length overtime. These are just some critical points to consider. If you are working closely with a mentor in your lab, I would suggest talking to your mentor about these questions and try to come up with ways to address and/or resolve them.

Feel free to post back with more questions!

Connie

[SciB]

Hi Roses,

I see Connie posted a similar set of questions to the ones I asked and you still did not answer them. We cannot help you to do a good science project if you do not tell us your hypothesis and plans for experiments.

Are you calling companies or just emailing? You will get a much better response if you talk to a human directly. Tell them that you are a high school student trying to do a great science fair project but need help to get the reagents to do the tests. They are more likely to help you out if you are on the phone with them.

I did a search for "cheap cortisol test kit" and found one for about $200, but I'm sure they would give you a student discount if you talked to them:

http://www.arborassays.com/product/k003 ... l-eia-kit/

Good luck,

Sybee

[deleted-132180]

Hi theroses,

This study just came out about how different environmental stresses can alter telomere length. Perhaps it may be useful for you in trying to develop a question, hypothesis, and experiments!

http://www.plosgenetics.org/article/inf ... en.1003721

Cheers,
Connie