I am working in bone regenerative technology and am well aware of Ca ion staining assay known as Alizarin red staining. This assay is said to work by binding to calcium ions in the cell, staining them red and later can be extracted by acetic acid (the red stain) which is subsequently studied by UV/Vis absorbance at 405 nm. Except, there are no definitive mechanisms explained in any literature on how it works, its specificity and the importance of its critical pH adjustment.
Dear Pranavkumar Shadamarshan, honestly asked:
did you really search or at least search enough for ancillary literature/papers?
At least in CONN's Biological Stains, 10th edition (HOROBIN R.W. and KIERNAN J.A., Eds, BIOS,Scientific Publishers Ltd, OXFORD 2002) I see some older references (1985,1991,1999,2000,2002,which might (eventually) contain valuable information on your request.
Citation from CONN's 10th ed. p.370/371:
"Metal complexation behaviour: forms complexes with a variety of metal ions, including aluminium and calcium. .... The hydroxy substituted anthraquinone chromophore can chelate metal ions; a sulfonate constituent is also present. Overall size of the hydrophilic anion is small, as is the conjugated system.... "
NB: Honestly: what about searching more efficiently? : e.g., I within some second found: http://www.sciencedirect.com/science/article/pii/S0021979702001571 or http://journals.sagepub.com/doi/abs/10.1177/17.2.110 and even more papers to dig into....'mechanism of staining' .... Good luck and best regards, WM.
Dear Pranavkumar Shadamarshan, honestly asked:
did you really search or at least search enough for ancillary literature/papers?
At least in CONN's Biological Stains, 10th edition (HOROBIN R.W. and KIERNAN J.A., Eds, BIOS,Scientific Publishers Ltd, OXFORD 2002) I see some older references (1985,1991,1999,2000,2002,which might (eventually) contain valuable information on your request.
Citation from CONN's 10th ed. p.370/371:
"Metal complexation behaviour: forms complexes with a variety of metal ions, including aluminium and calcium. .... The hydroxy substituted anthraquinone chromophore can chelate metal ions; a sulfonate constituent is also present. Overall size of the hydrophilic anion is small, as is the conjugated system.... "
NB: Honestly: what about searching more efficiently? : e.g., I within some second found: http://www.sciencedirect.com/science/article/pii/S0021979702001571 or http://journals.sagepub.com/doi/abs/10.1177/17.2.110 and even more papers to dig into....'mechanism of staining' .... Good luck and best regards, WM.
Note added: It might also be essential, to have a look into the huge amount (100+) of Questions and Answers (Q/A) to be found already in the ResearchGate Archive,.. you only have to SEARCH - (in) QUESTIONS (for) ALIZARIN (creates an URL: https://www.researchgate.net/search.Search.html?type=question&query=ALIZARIN ), where somebody might have posted already the answer to your question(s).
The most interesting "old" paper might be the article by PUCHTLER H. et al, 1969: ON THE HISTORY AND MECHANISM OF ALIZARIN AND ALIZARIN RED S STAINS FOR CALCIUM.
@Wolfgang H. Muss Thank you very much, sir. Helped me a lot. I could not find an effective explanation in the papers that I went through and I did not know how to search for it. Your help is much appreciated.
Dear Pranavkumar Shadamarshan , you're welcome.
Best regards, W.H.M.
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