Hi,
I have mistakenly added tissues into the Trizol reagent and snap froze it. Later I realized I did a mistake and immediately removed the tissues out of Trizol and rinsed with PBS.
Does it make any difference while we Western Blotting. I think it should not make any hindrance.
Any expert suggestions please.
Regards,
Harsha
Hi! well, if I was you I will try to lysate and extract the proteins of two samples, run the acrylamide gel, do the transfer to the blotting membrane and stain with Ponceau S (red) in order to see if there is any band (protein) from your samples. In one morning you may know something.
good luck!
I would suggest to use another tissu as a control and let it to stay in Trizol as you did for your valuable sample. After, you extract the proteins and analyse them as Isabel told you you can proceed with you sample or not. In the case you do not see proteins I may suggest to proceed with your sample as suggested by Trizol protocol to extract RNA, DNA and proteins.
Thank you Laurence Stuart Dawkins-Hall Isabel Forner-Piquer and Abderrahmane Bengrine for your inputs.
As we have samples which were not put in Trizol, we'll do comparison among those tissues. Lets see.
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