I want to use them to measure SERCA pump activity in mice skeletal muscles.
Hello,
First you have to calculate the composition of your solution using the software computer program based on the work of Fabiato (Fabiato, A. & Fabiato, F. Calculator programs for computing the composition of the solutions containing multiple metals and ligands for experiments in skinned muscle cells. J. Physiol. 75, 463-505)
You will download references and software at this web page : http://maxchelator.stanford.edu/references.htm
You can also see the following references, were the authors performed some studies in those they calculated pCa for their solutions :
Calculated and measured [Ca2+] in buffers used to calibrate Ca2+
macroelectrodes John A.S. McGuigan, Friederike Stumpff Analytical biochemistry (2013) 436 : 29-35
Analysing force–pCa curves John S. Walker, Xiaotao Li and
Peter M. Buttrick Muscle Res Cell Motil (2010) July ; 31(1): 59–69
Finally I advice you to use stock calcium solution already prepared (for example CaCl2 1M from Sigma Ref 21115-100ML) instead of preparing a self-made solution with calcium chloride powder. As calcium hydrates, it is very complicated to weight correctly and to have the good final concentration.
Good luck
Regards
Jean-Pierre Gomez
Dear Jeane Pierre. I did this sometime ago. Go to my Research Gate account and copy a book "Practical Mitochondriology". In the Chapter on Calcium there is a description and reference for the method.. Also, if you use ruthenium red, you must recristallise it..
Dear Jean and Alexander, thanks for your answers. I finally figured it out with the computer program and scientific calculator. :)
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