western blot analysis!

A

Hello any lab scientists out there!
I have been putting western blot results together for one of my pathetic looking chapters! I have not really done much WB except for these 6 gels at the very end of my lab work, and would really like to be able to do some quantitative analysis on them.

I was told there was free software out there and found ncbi's image, which unfortunately won't work with my lovely new mac! I have downloaded imageJ, however it seems impossible to use with no real instructions on how to use it :(

Does anyone know of this software or any other open source software that would do this nice little job for me and help me put some extra data and stats into my chapter :)

thanks to anyone who can help and save me hours of trawling and downloading!
:p

S

hi alpacalover
I'm not sure what you want to do, what exactly do you want to put in your thesis? Are you looking for 3-D images for your band from your gel? what kind of data or stats are you looking for?
cannot help if we dont know what you want.
satchi

A

Hi Satchi,
doh!! apologies for being an idiot - really shouldnt do these things later at night when i'm shattered! :)

I was hoping to be able to use software to get the relative densities of each band so that I can compare them better, get some actual numbers and not "this band is bigger and darker than that band...." etc!
does that make sense? Not really good with westerns, i'm a microbiologist so not really my area!

Cheers

S

hi alpacalover,
to quantify the "brightness" of your band (how intense it is) instead of visual comparison with ladders then you need something like Genetools software. http://www.genegnome.com/html/products.html
Or gelQuant http://www.ampl.com.au/gq_frames.htm.

I haven't used open source applications for this. There are lots of other free apps but these are mostly for gene sequencing. http://molbiol-tools.ca/molecular_biology_freeware.htm

What you need to do is to go to the molecular labs in your university, ask the students there what software they are using to quantify their bands, and ask if you can have access to it. If you need to get permission from their supervisors, then ask first.

You only have 6 gels, this will be very fast. I don't think anyone will turn you down. Usually people in labs help each other. Ask someone to help you, make friends and ask for a suitable time so that this person may show you how to do it for one gel, and then you can do the other 5 gels. Its not like you have 600 gels.

Usually the software will be in the computer near the camera system used to photograph the gels. Genetools can be run on a mac but as I said, if your university only uses PCs, it is better to run the analysis on a lab in the computer and then just use the data on your mac.

Molecular labs usually will have this kind of software. Go and ask tomorrow. good luck! :-)
regards
satchi

A

HI Satchi!

Thank you very much for advice, I will try and get hold of someone at uni, i've got a few friends who do them alot so i'm sure someone would be able to help. Its just that i'm now not working at uni and full time else where therefore its a bit more difficult.

But for the sake of afew gels it really would be stupid to try and get software and learn it myself. I have a friend that has just submitted so i'm sure he'd be willing to take some time to help me :)

Cheers, AL

S

Hi there,

I use Image J to quantify western blots. It is very easy to use once you know how. I don't have much time to explain now but if you have msn messenger or pm me an email addresss I can forward some instructions on how to use it. I have a spreadsheet with some old data in which you might find useful. It shows all the steps of how to make it into a graph.

Superman.

A

Thanks Superman, i've sent you a PM!

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