ECL and detection in western blot?

Chinkyu Lee @Chinkyu_Lee

07 July 2016 2 8K Report

Hello,

Just few days ago, I asked almost same question about unwanted mixing of ECL during western blot

and I got great helps from replies. I appreciate those who responded again.

But I think that I described my previous question a bit ambiguously.

To sum up the answers,

The answeres replied that 'ECL remained in unchanged vinyl' would not

affect the net result of 'further detections after quick washing ' because the

volume remained is minimal.

I forgot to refer to 'absence of wiping of previous ECL' in unchanged vinyl.

Of course, after wiping, the amount of previous ECL would be minute,

but i wanted to ask about the case when I forgot to wipe the vinyl

to minimize previous ECL for further detections after quick washing.

Currently i am more cautious and professional about western blot so I change transparent vinyl every time for later detections even though in one particular western blot experiment.

But I am concerned whether I can believe my seemingly reliable detection

results obtained in the past, when I can't remember whether I wiped ECL

or exchanged with new, clean vinyl,

considering

the 'unwanted effect of the remained ECL 'on 'further detections after quick wahsing'

caused by

absence of wiping or absence of vinyl exchange without wiping remained

ECL.

And lastly, wiping cleanly whole casette before further detections would be

good, but absence of wiping whole casette may affect the results?

I want to get opinions about my concern above.

Thank you!

Nancy Duarte

Well, I actually prefer use a new piece of plastic everytime I am going to perform the chemiluminiscent detection because it increases the reliability of the results, since you are using new material that hasn't been involved before in a previous detection procedure. However, as you mentioned before, the remaining reagent is just a little amount, so that I think that your results won't be really affected if you forgot to change the piece of plastic before.

Chinkyu Lee

Hello Nancy Duarte,

Thank you so much for your great answer.

I learned good things from your help.

I appreciate again.

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