I want to measure, whether there is one compound in the extra-cellular matrix. I know there is plenty of it inside cells. How shall I proceed to be sure I do not break the cells?
If I crush the material in ball mill and extract with buffer without detergent, that will not ensure there is nothing from inside of the cell, right?
You are correct, crushing will get inside as well as out. But you seem to be well-versed in plant tissue culture so if you created a protoplast you could compare levels from the cells versus the enzymatic mess that includes everything that was outside the cell. But if there is plenty inside, can you be sure that no protoplasts pop open because that would really skew your result.
A little more information about what you hope to find would help. Is it a chromophore, or does it fluoresce if hit with the proper wavelength? If so, other options might be found. Is this compound a peptide or secondary chemical?
Hi, thanks for your answer. Personally I'm not so experienced with plant tissue culture, I have never done protoplast, but some colleagues do.
I wish to examine, whether there is some extracellular ATP. The "easiest" detection would be probably on HPLC-UV/VIS.
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