Hi Harpreet,

did you try this protein analyzer? It was made by one of my collaborator and is really reliable?

http://image.bio.methods.free.fr/ImageJ/?Protein-Array-Analyzer-for-ImageJ,25

Hope it helps,

Sarah

Hi,

Measure the density of the spots with a software like Image J. Substact the background of all your spots (background = density of the spot of your negative control). Normalize every spots with your spots "positive control" for each membrane.

After these transformations of your spots you can make a ratio of your "spots experimental tests"/"spots experimental control" to express a change in the expression of the angiogenic factors.

Let me know if it is not clear enough.

Good luck.

William

Hi,

You can follow these steps.

1) Measure the pixel density of each spot with Fiji or image J 

2) Subtract  the background.

3) Normalize all the values with reference spot(s) values on the blot.

4) I think with the kit they will provide you a transparent sheet with each spot labelled accordingly. You can check on the booklet provided with a kit which spot corresponds to what.

5) Compare your treated sample with the untreated. 

Thats it.

I hope it is clear for you.

Good luck..

Jagadeesh

Many Thanks for your help William and Jagadeesh.

When you say normalize all the values with the reference spot values on the blot, what exactly do you mean?

Many Thanks,

Harpreet

Hi everyone,

Please could you help me to understand which steps I need to follow in Image J to do the analysis of my proteome array ?

Thank you very much.

Could anybody provide or recommend publications that have used this array? I would like to know what are the minimum pixel values that should be interpreted as significant biological levels of secreted angiogenic factors.

In our assays, we found values ranging from 54 to 80,000 pixels.

Thank you!