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Ponceau S dilemma


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#1 Adrian K

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Posted 27 October 2010 - 07:55 PM

Hi all,
I had come across few protocols of ponceau S stain of pvdf membrane.
Some uses "Trichloroacetic" and some uses "Trifluoroacetic" acid.

Which is better / more relevant? Also, can share your formulation of "ponceau S"?

thanks a billion.
Expecting the world to treat you fairly because you are a good person is like expecting the lion not to attack you because you are a vegetarian.

..."best of our knowledge, as far as we know this had never been reported before, though I can't possible read all the published journals on earth, but by perform thorough search in google, the keywords did not match any documents"...

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#2 biotef

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Posted 28 October 2010 - 06:55 PM

Hi all,
I had come across few protocols of ponceau S stain of pvdf membrane.
Some uses "Trichloroacetic" and some uses "Trifluoroacetic" acid.

Which is better / more relevant? Also, can share your formulation of "ponceau S"?

thanks a billion.



I assume you just want to check efficiency of transfer and then go for immunoblotting. What we use is 0.1% Ponceau S in 5% Acetic acid [Not trifluoroacetic acid] Stain for 5-10 mins. rinse once or twice with water. mark the marker bands and then destain using 0.1 M NaOH 2-3 mins followed by subsequent rinsing with water 2-3 times / 5 mins to remove NaOH. then perform blocking and primary antibody treatment. hope this helps you.

#3 Adrian K

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Posted 29 October 2010 - 09:13 AM

Hi biotef,
Thanks for your reply and protocol sharing. The reason I ask is because all the links listed below uses either one of the acid that I mentioned.. that is the reason I find it curious...

Thank you very much
Adrian



Article: http://www.scielo.br...ipt=sci_arttext
"The nitrocelullose membrane was revealed with 0.2 % ponceau S in 3 % trifluoroacetic acid (v/v) and destained with PBS"

Milipore: http://www.millipore...ottingprotocols
"Stock solution: 2% Ponceau S in 30% trichloroacetic acid and 30% sulfosalicylic acid; dilute 1:10 for use."

Ivan: http://ivaan.com/protocols/137.html
"Mix 2 g of Ponceau-S (2%), 30 g of Trichloroacetic acid (30%), 30 g of Sulfosalicylic acid (30%), and dH2O to 100 mL"
Expecting the world to treat you fairly because you are a good person is like expecting the lion not to attack you because you are a vegetarian.

..."best of our knowledge, as far as we know this had never been reported before, though I can't possible read all the published journals on earth, but by perform thorough search in google, the keywords did not match any documents"...

"what doesn't kill you, makes you stronger"---Goddess Casandra reminds me to be strong

"It's all just DNA. Do it."---phage434




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