Difference between revisions of "Talk:Respirometry"
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== MiPNet discussion forum: lymphocytes and respiratory measurements (2013-10-24) == | == MiPNet discussion forum: lymphocytes and respiratory measurements (2013-10-24) == | ||
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Lymphoblasts have been examined in here: http://www.ncbi.nlm.nih.gov/pubmed/22427588, and more recently here: http://bioblast.at/index.php/Pecina_2013_Abstract_MiP2013. Generally, for experiments in permeabilized cells [you do not (and would not want to) permeabilize the mitochondrial membrane with digitonin, but the plasma membrane], you need to optimize conditions; I suggest you follow the protocol given in this reference: http://bioblast.at/index.php/Pesta_2012_Methods_Mol_Biol (download "Bioblast pdf"). | Lymphoblasts have been examined in here: http://www.ncbi.nlm.nih.gov/pubmed/22427588, and more recently here: http://bioblast.at/index.php/Pecina_2013_Abstract_MiP2013. Generally, for experiments in permeabilized cells [you do not (and would not want to) permeabilize the mitochondrial membrane with digitonin, but the plasma membrane], you need to optimize conditions; I suggest you follow the protocol given in this reference: http://bioblast.at/index.php/Pesta_2012_Methods_Mol_Biol (download "Bioblast pdf"). | ||
Further, there is a recent paper (using our competitor's equipment - but, so what?) examining respiration in INTACT blood cells (platelets, lymphocytes, monocytes, neutrophils), at least this should be easily possible with an oxygraph (http://www.ncbi.nlm.nih.gov/pubmed/23528848). | Further, there is a recent paper (using our competitor's equipment - but, so what?) examining respiration in INTACT blood cells (platelets, lymphocytes, monocytes, neutrophils), at least this should be easily possible with an oxygraph (http://www.ncbi.nlm.nih.gov/pubmed/23528848). | ||
Revision as of 10:30, 24 October 2013
MiPNet discussion forum: lymphocytes and respiratory measurements (2013-10-24)
Magda Labieniec-Watala
Lymphocytes and respiratory measurements - can anyone help? I have just started to work with lymphocytes isolated from blood and I am interested in measuring their respiratory capacity using oxygraph. I have used the MIRO6 medium but probably it was a mistake. Lymphocytes do not respire and do not respond to substrates and inhibitors in this environment. Thus, I have a request for experimental protocol for these measurements from those who study the mitochondrial respiratory in lymphocytes. May enyone do similar experiments and have an appopriate experience? I would like to add that we do not isolate mitochondria from lymphoctes but we use digitonin in order to permeabilize the mito membrane. I tried to find some papers concerning these studies, but without success. I would be very grateful for some suggestions in this issue. Thank you in advance. [[email protected]]
Gerhard Krumschnabel
Lymphoblasts have been examined in here: http://www.ncbi.nlm.nih.gov/pubmed/22427588, and more recently here: http://bioblast.at/index.php/Pecina_2013_Abstract_MiP2013. Generally, for experiments in permeabilized cells [you do not (and would not want to) permeabilize the mitochondrial membrane with digitonin, but the plasma membrane], you need to optimize conditions; I suggest you follow the protocol given in this reference: http://bioblast.at/index.php/Pesta_2012_Methods_Mol_Biol (download "Bioblast pdf"). Further, there is a recent paper (using our competitor's equipment - but, so what?) examining respiration in INTACT blood cells (platelets, lymphocytes, monocytes, neutrophils), at least this should be easily possible with an oxygraph (http://www.ncbi.nlm.nih.gov/pubmed/23528848).
