Cookies help us deliver our services. By using our services, you agree to our use of cookies. More information

MiPNet10.04 CellRespiration

From Bioblast
Publications in the MiPMap
Garedew A, Haffner B, Huetter E, Gnaiger E (2005) An experiment with high-resolution respirometry: Phosphorylation control in cell respiration. Mitochondr Physiol Network 10.04.

» Open Access-MiPNet10.04.pdf; Versions

O2k-Protocols contents

OROBOROS (2005) Mitochondr Physiol Network

Abstract: O2k-Protocol for Phosphorylation Control in Cell Respiration

Methodological and conceptual features of highresolution respirometry are illustrated in an experiment with cultured, suspended cells in the OROBOROS Oxygraph-2k (O2k). The experiment demonstrates manual titrations of inhibitors, and automatic titrations of an uncoupler using the electronic Titration-Injection microPump TIP2k . Application of the DatLab 4.3 (upgraded) software is shown for instrumental control (O2k and TIP2k), and on-line data analysis . The following guideline describes the experiment in the form of a laboratory protocol, complementary to the relevant sections of the Oxygraph-2k Manual. The experiments were carried out by participants of an O2k-Course on high-resolution respirometry in April 2005 (IOC30; Schröcken, Austria).

>> O2k-Protocols:Overall contents
>> Product: OROBOROS Oxygraph-2k, O2k-Catalogue
Figure 1. Respiration of 32D cells (1.1106 ml-1). Traces for the two chambers of the Oxygraph-2k are superimposed. A: Oxygen flow [pmol O2s-1106]; B: Oxygen concentration [μM] (graph layout “7 Gr1-Flux Gr2-O2 Conc”). After inhibition of ATP synthase with oligomycin, FCCP was titrated at 0.5 μM steps with the Tip2k. Each titration step is automatically marked by an event (vertical lines). After the aerobic-anoxic transition, the chambers were opened for re-oxygenation, causing a 10-min disturbance of the traces of respiratory flow. The Tip2k needles were removed before opening, and re-inserted after closing the cham.


O2k-Network Lab: AT_Innsbruck_OROBOROS


Labels: MiParea: Respiration, Instruments;methods 


Organism: Mouse  Tissue;cell: Blood cells  Preparation: Intact cells 

Regulation: Coupling efficiency;uncoupling, Uncoupler  Coupling state: LEAK, ROUTINE, ETS"ETS" is not in the list (LEAK, ROUTINE, OXPHOS, ET) of allowed values for the "Coupling states" property. 

HRR: Oxygraph-2k, TIP2k, Protocol"Protocol" is not in the list (Oxygraph-2k, TIP2k, O2k-Fluorometer, pH, NO, TPP, Ca, O2k-Spectrophotometer, O2k-Manual, O2k-Protocol, ...) of allowed values for the "Instrument and method" property. 

O2k-Demo 

Further information

>> Excel Templates and DatLab-Demo Files
  • Version 6: 2012-04-07
  • Version 1: 2005