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| Ā | | [[Image:BB-Bioblast.jpg|left|40px|link=http://www.bioblast.at/index.php/Bioblast:About|Bioblast wiki]] |
| == Pmt, PC from literature? ==
| | == Popular Bioblast page == |
| '''Question:'''Ā Ā (1) With the TPP spreadsheets, Pmt and Pc values are the values that we use literature values for (that actually measured membrane content), and not actually measured protein (mg/mL) off the sample that is being run? and Vmt I think we also used published values for?
| | [[MiPNet15.08 TPP electrode]] has been accessed more than |
| Ā | | ::::* 10,000 times (2019-12-12) |
| '''Answer:'''Ā No, Pmt and Pc describe how much sample you put into your chamber andĀ therefore can not be taken from literature. If you use twice the amountĀ of sample these values will have to double. For isolated mitochondriaĀ Pmt = Pc.
| | ::::* 5,000 times (2016-02-04) |
| Please see also the Rottenberg paper cited.
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| Vmt can be taken from literature, especially as the TPP method is very insensitive against errors in Vmt.
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| Similarly we take Kin and Kout from literature. Vmt, Kin , and Kout are all SPECIFIC values, that is values per amount of mitochondrial /Ā cellular protein.Ā But to use these specific values you needĀ theĀ absolute values of mitochondrial cellular protein- ideally from anĀ assay or at least some rough estimation from the respiration itself (ifĀ you know the typical respiration of your sample per mg mitochondrialĀ protein). In the sheet "external input" you see a very rough templateĀ for this respiration approach, but a proper assay is of course the wayĀ to go.
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| SomeĀ background information :-) : Actually the choice ofĀ mitochondrial protein as the marker the specific valuesĀ Kin andĀ KoutĀ is a bit arbitrary /Ā counter intuitive: The unspecific binding, described by Kin and Kout, actually will take place mainly in theĀ mitochondrial lipids not in the proteins. But since mitochondrialĀ protein is easy to measure and the ratio lipid to protein is hopefullyĀ constant all the initial developers of the method used mitochondrialĀ protein as a marker for the amount of mitochondria. For samples beyondĀ isolated mitochondria (permeabilized cells,..) the assumption of aĀ constant lipid to protein ration of course brakes down, maybe one reasonĀ why there are no good values for Kout in "cellular samples". [[User:Fasching Mario|Fasching Mario]] 09:41, 17 April 2015 (CEST)
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