SUIT-001 O2 ce-pce D004

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SUIT-001 O2 ce-pce D004



Abbreviation: RP1 ce-pce blood

Reference: A - SUIT-001

SUIT number: D004_ce1;1Dig;1PM;2D;2c;3U;4G;5S;(6Oct);7Rot;8Gp;9Ama;10AsTm;11Azd

O2k-Application: O2

MitoPedia: SUIT - SUIT reference protocol Reference protocol RP1 for for living PBMC and PLT cells (ce, ROUTINE respiration) and cells that are permeabilized in the chamber (pce)
SUIT protocol pattern: 1PM;2D;2c;3U;4G;5S;(6Oct);7Rot;8Gp-

The SUIT-001 O2 ce-pce D004 protocol is the reference protocol 1 (RP1) specifically for PBMCs and PLTs. The main difference between the RP1 for PBMCs and PLTs and RP1 for other mitochondrial preparations ( see SUIT-001) is the absence of Oct in the case of blood cells. The SUIT-001 O2 ce-pce D004 protocol in combination with SUIT-002 O2 ce-pce D007a is specially designed to provide a common reference for comparison of respiratory control of PBMC and platelets cells (non-permeabilized cells must be added to the chambers and then permeabilized as a part of the protocol) in a wide variety of species. SUIT-001 O2 ce-pce D004 gives information of ROUTINE respiration (ce: non-permeabilized cells) and after the plasma membrane permeabilization (pce: permeabilized cells) of the linear coupling control (L- P- E) with NADH linked-substrates (PM). Moreover, the pathway control in ET state (N, NS, S and SGp pathways) can be evaluated by using this SUIT protocol. SUIT-001 O2 ce-pce D004 can be extended with the CIV assay module.

Communicated by Doerrier C and Gnaiger E (last update 2019-09-24)

Representative traces

D004 O2 traces.png

MitoPedia: SUIT

Steps and respiratory states

SUIT-001 for PBMCs and PLTs

Step State Pathway Q-junction Comment - Events (E) and Marks (M)
ce1 ROUTINE ce1
  • ROUTINE respiration in the physiological coupling state R. Externally added permeable substrates could contribute to this respiratory state.
1Dig ROX ce1;1Dig
  • Optimum effective digitonin concentration for complete plasma membrane permeabilization.
Step State Pathway Q-junction Comment - Events (E) and Marks (M)
1PM PML(n) N CI PML or PM_L: Pyruvate & Malate, N-LEAK respiration, NL
2c PMcP N CI 1PM;2D;2c
  • NADH-linked substrates (type N-pathway to Q).
  • OXPHOS capacity, P (with saturating [ADP]), active OXPHOS state.
  • Addition of cytochrome c yields a test for integrity of the mtOM. Stimulation by added cytochrome c would indicate an injury of the mtOM and limitation of respiration in the preceding state without added c due to loss of cytochrome c. Typically, cytochrome c is added immediately after the earliest ADP-activation step (P, OXPHOS capacity with saturating [ADP]).
(6Oct) 1PM;2D;3U;4G;5S;(6Oct)
  • Oct is skipped in PBMCs and PTLs
7Rot SE S CII 1PM;2D;3U;4G;5S;(6Oct);7Rot
8Gp SGpE SGp CII&GpDH 1PM;2D;3U;4G;5S;(6Oct);7Rot;8Gp
  • Respiratory stimulation by action of succinate and glycerophosphate, Gp, with convergent electron flow in the SGp-pathway (CII&GpDH-linked pathway to the Q-junction).
  • Noncoupled electron transfer state, ET-state, with ET capacity, E.
9Ama ROX
  • Rox is the residual oxygen consumption in the ROX state, due to oxidative side reactions, estimated after addition of antimycin A (inhibitor of CIII). Rox is subtracted from oxygen flux as a baseline for all respiratory states, to obtain mitochondrial respiration (mt).
Step Respiratory state Pathway control ET-Complex entry into Q-junction Comment
## Azd ROX


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Strengths and limitations

  • SUIT-001 in combination with SUIT-002 provides a common reference for comparison of respiratory control in a large variety of species, tissues and cell types. Both SUIT protocols provide a mitochondrial mapping which allows:
1. to obtain reference values.
2. to evaluate mitochondrial physiological diversity, generating a mt-database on comparative mitochondrial physiology.
3. to screen specific defects.
  • SUIT-001 and SUIT-002 are used in the MitoFit Proficiency test for inter-individual and inter-laboratory reproducibility quality control.
  • A succinate concentration of >10 mM may be required for saturating SE capacity.
+ Linear coupling control (L-P-E) with N substrates (PM).
+ Pathway control in ET state (N, NS, FNS, S and SGp pathways).
+ Harmonization with SUIT-002 allows to perform both SUIT protocols in parallel. The cross-linked respiratory states can be statistically used as repeated measurements.
+ In blood cells, SUIT-001 and SUIT-002 harmonize in: ROUTINE (ce1), SGpE (8Gp in SUIT-001 and 10Rot in SUIT-002) and Complex IV.
+ Harmonization with many SUIT protocols (up to step 5S).
+ This protocol can be extended with the Complex IV module.
+ The addition of non-permeabilized cells to the chambers provides additional information (ROUTINE respiration) which can not be obtained in SUIT-001 O2 mt D001 where the cells added to the chambers are already permeabilized.
- The substrate combination for maximum ET capacity (FNSGP) is not obtained in SUIT-001 in favour of measuring SE.
- Oct is not added in ET state.
- Lengthy duration of the experiment.

Compare SUIT protocols

  • SUIT-001 O2 ce-pce D004 (SUIT-001 for permeabilized PBMC and PLT) differs from SUIT-001 O2 ce-pce D003 (general SUIT-001 for permeabilized cells) in the absence and presence of octanoylcarnitine, respectively.
  • SUIT-002 O2 ce-pce D007a (RP2): Harmonized SUIT protocol for PBMC and PLT cells (non-permeabilized cells must be added to the chambers and then permeabilized as a part of the protocol).
  • SUIT-001 O2 mt D001: SUIT-001 for permeabilized cells (cells added to the chamber have been permeabilized before their addition). This SUIT protocol provides no information on ROUTINE respiration of non-permeabilized cells (ce).
  • SUIT-004: Short version of RP1; without G, F and Gp.
  • SUIT-006: Very short initial coupling control module.


MiPNet21.06 SUIT RP2018-06-25
SUIT reference protocol for OXPHOS analysis by high-resolution respirometry.
Doerrier 2018 Methods Mol Biol2018Doerrier C, Garcia-Souza LF, Krumschnabel G, Wohlfarter Y, Mészáros AT, Gnaiger E (2018) High-Resolution FluoRespirometry and OXPHOS protocols for human cells, permeabilized fibers from small biopsies of muscle, and isolated mitochondria. Methods Mol Biol 1782:31-70.Human
Saccharomyces cerevisiae
Skeletal muscle
Endothelial;epithelial;mesothelial cell
Blood cells

MitoPedia concepts: SUIT protocol, SUIT A, Find 

MitoPedia methods: Respirometry