Hello,
I always get great support and help from Researchgate,
And i am sorry to anyone whom i forgot to express my gratitude for reply.
Some days ago, i asked same question about sample buffer of Western blot.
I use 4XSB as a sample buffer and i use this buffer after it melts.
(I store the buffer at -20, not room temperature)
I always quantify protein concentration and make same concentration of proteins between all of my samples for reliable research. It is ok, but
I want to ask whether absence of vortexing or tapping sample buffer
after it melts is critical to results.
Can i conclude that if no problems found, absence of vortexing or tapping
my buffer described above does not affect research outcomes critically?
It has been just short time for me to begin research in my field, so
even though i get results seemingly without no problems, i think it is
important for me to get feedback or comments related to procedures
from more advanced researchers.
Thank you very much!
Hey Lee,
For some samples it won't be any problem if you mix your Buffer prior adding or not. But I would do it anyway, just to be sure, that the buffer is mixed correctly. It can happen, that the salts in your buffer melt slower than water or your denaturing agent.
So even if most of the time you wont see a difference, i would mix it anyway, so you wont need to worry.
Cheers!
Hi
I agree with Simon and all what he right I will add that the SDS present in some sampling buffers precipitate at low temperature and it need to be heated and mixed well before been used. The fact that you didn't face any problem yet it does not mean that you are not going to have some in the future. So please do it
Dear Simon
Thank you for your informative and kind reply!
So you mean that good results with no mixing of sample buffer
means that absence of mixing sample buffer melted does not act as
Critical problem? But nevertheless, mixing sample buffer is
definitely optimal procedure?
Thank you again.
Dear Abderrahmane,
I appreciate your response!
So you mean that results without problems in spite of absence of
mixing sample buffer are reliable? But nevertheless, mixing the buffer
is optimal and can affect results critically some day?
Thank you for your kindness again
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