Hi guys,
I've been asked to source all the equipment and reagents needed to set up western blot in the lab, starting from scratch (all we have is a Nupage electrophoresis tank). I know my preferences for transfer system, membranes, reagents.... but I'm having doubts on the developing method. In the past I've always used ECL and x-ray films, but I know more and more labs these days favour other chemiluminescent reagents and imaging as you get a better control on exposure and a better dynamic range.

So here's my question.

What method do you use to develop a western?


Please share your opinions

Thanks!

Films still have better resolution and usually sensitivty than cameras in my experience. However, the cost of running a darkroom with film and developing chemicals and associated equipment is quite high. Modern cameras are pretty good though and give nice images with reasonable exposure times (compared with 3-5 years ago) based on chemiluminescent detection.

Have you thought about chromogenic detection?

I"ve used both X-rays and digital imaging- we have a Fujifilm CCD system in the lab now, which is very convient- less mess and less time for developing as our nearest dark room in 2 floors below us and we have to book in. The advantage of the digital images is that they are easily transferred and you can carry out some quantitative analysis on them more easily too.
I agree the sensitivity of the films can sometimes be better, but its not been an issue for me. I'm pretty converted to the Digital imaging now.
We use in conjunction with ECL, but our system allows us to take images in a number of wavelengths so can be used for other methods too.

Thank you guys! putting a list together now, will see what my manager goes for

I would almost always go for the CCD variant - you always get the best exposure that way.

But beware, there is a HUGE difference between imagers - some are really crap, some are really good, and the price alone often doesn't tell you which is which.

Thanks for pointing that out. What do you think about Alpha Innotech? My old lab now has that and they are quite happy. Which one would you recommend? As you say there are many and I'm no expert in imagin so I dont even know what I have to look for.

I have used an older Alpha Innotech, that was a good one, the current ones from Bio-Rad also aren't bad.
We settled on the MF-ChemiBIS which was the best deal overall (although the software has its issues sometimes).

Hey
We've been working with films the last years, but we are changing to a camera now (cleaner and handier). We asked 3 companies to show their equipment: we had fujifilm (nice and simple, bit limited), bertholds (nope bad experience, representatives didn't manage to make a good picture) and Westburg (nice! and a good price )
Hope this can help you?
Grs

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we are changing to a camera now
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so what is the problem in that.....you can go through different sites and books and enhance your knowledge in this field...