Hi,
I wonder if there is any buffer that i can keep tissues in, right after the dissection (mice). That would be very handy because there will be a lot of sample collection going on during dissection.
Later on we will homogenize them for ELISA.
I keep the ones that will be used for RNA isolation in RNAlater but i dont know any for ELISA. Would the RIPA Lysis and Extraction Buffer be a choice?
Any experience on that?
Thanks!
Hi Bursa
I think the best alternative would be storing your tissue in RNAlater®
RNAlater treated samples can be stored at room temperature (25°C) for up to one week, at 4°C for up to one month, or at -20°C (or lower) indefinitely. Intact protein can be recovered from RNAlater-stabilized samples for use in downstream applications such as Western blotting or 2D gel analysis.
https://www.thermofisher.com/in/en/home/references/ambion-tech-support/rna-isolation/tech-notes/rnalater-for-rna-and-protein-stabilization.html
Thanks @ Ashis Kumar
I think RNA later denatures proteins. Seems like it would be ok for western blotting but wouldnt that be a problem when running ELISA?
Hi Bursa!
I totally agree with you!
Find out if the antibodies used in your ELISA experiment, can also be used for other assays, like Western blot or IHC. If Yes, the chances are high, that your Antigen in its denatured form because of RNAlater would still be detected. If not - ELISA antibodies directed against native epitopes or modified epitopes - then the chances are not that good.
I have used RIPA buffer for ELISA and encountered problems in coating! could be due to the effect of detergent.
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