Hi guys,
From my expirence there are pros and cons of with or without plastic sheet cover:
1. Plastic sheet cover sheet is good to keep the blot wet, good for long time exposure, but it will have some noise/blur background from the sheet, even with clean sheet (compare to non-cover).
2. Non-cover blot will have no noise/background from thecover sheet, however the blot can be dried during the exposure, and have to be careful to place the blot to avoid bubble between tray and the blot.
I haven't tried plastic wrap yet, but I expect wrickle with it.
Does it have any solution to cover both pros of these method? So the It can keep blot wet without create background from the cover.
Thank you
Cheerio,
Hi Asa,
I used to do the western blot by placing the nitrocellulose membrane between saran wrap. By this point I would use a clean tissue to remove any air bubble and striating the wrap to become wrinkle free.
If I ever had a high background noise, I always knew it was not from the wrapping and I would proceed with high salt wash to remove persistent background before placing into a saran wrap. If you like I can give you a short protocol for salt wash.
Good luck,
Ali
Hi Asa,
I learned in a lab a nice way to overcome that nois and keep the blot wet enough:
The last step, where you put the solution on your blot for detection - do it in a culture dish. Put your blot in the right way and after it was long enough in that solution you can flip the dish (the solution shouldn't be in that culture dish anymore at that point). Your blot will stick to the culture dish without any bubbles, because there is a thin layer of the solution remaining. You can your culture dish with the blot in that way beneath your CCD camera and can shot nice pictures.
Look for the pdf file I attached. It shows in an clip-art, what I just described you.
Have fun,
Mandy
when I am using a ccd digital camera I will plate the blot between two very thing glass pieces. This helps to keep the blot from drying out and helps to reduce the refraction. You want to use as thin of pieces of glass as you can find as thicker glass refracts light more.
We have successfully used "sheet protecters" when photographing blots. They have to be perfectly clear thin ones to work well. They are MUCH easier to use than plastic wrap for removing/avoiding bubbles both below and above the blot and maybe easier than using glass since they are both flat AND flexible. Don't try to slip the blot into the protector, but instead cut the protector on three sides leaving the front and back connected on one side. On a dry surface carefully place the plot on the 'open' protector sheet pushing out any air bubbles. Then slowly lower the top from the connected side to the open side again pushing out bubbles. The blot can be stored between the protector sheets taking up less space than folded plastic wrap. Try it and let me know if it didn't work for you!
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