Carboxyatractyloside: Difference between revisions
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::::* Titration volume: 1 ยตL using a 10 ยตL Hamilton syringe (2 mL O2k-chamber). | ::::* Titration volume: 1 ยตL using a 10 ยตL Hamilton syringe (2 mL O2k-chamber). | ||
::::* Final | ::::* Final concentration: 1 ยตM. | ||
Revision as of 14:24, 27 August 2020
Description
Carboxyatractyloside, Cat, is a highly selective and potent inhibitor of the adenine nucleotide translocator (ANT). Cat stabilizes the nucleoside binding site of ANT on the cytoplasmic (positive) side of the inner membrane and blocks the exchange of matrix ATP and cytoplasmic ADP. It causes stabilization of the c conformation of ANT leading to permeability transition pore (PTP) opening, loss of mitochondrial membrane potential, and apoptosis.
Abbreviation: Cat
Reference: MiPNet03.02
MitoPedia topics: Inhibitor
Application in HRR
- Cat: Carboxyatractyloside (carboxyatractyloside potassium salt from Xanthium sibiricum, C31H44O18S2*Kx); Sigma C 4992, 2 mg, store at -20 ยฐC; FW = 802.99 (free acid basis); Alternative source: Merck 216201-5MG (Calbiochem, Carboxyatractyloside potassium salt from Xanthium sibiricum, C31H43O18S2K3*3H2O), 5 mg, store at -20 ยฐC, FW = 939.1).
- Caution: Chemicals stored in the fridge or freezer should be allowed to reach room temperature before opening. Very toxic!
- Preparation of 5 mM stock solution (dissolved in H2O):
- Weigh 4.7 mg carboxyatractyloside.
- Dissolve in 1 mL H2O.
- Divide into 0.2 mL aliquots.
- Store at -20 ยฐC.
- O2k manual titrations ยป MiPNet09.12 O2k-Titrations
- Titration volume: 2 ยตL using a 10 ยตL Hamilton syringe (2 mL O2k-chamber).
- Final concenteration: 5 ยตM.
- Preparation of 2 mM stock solution (dissolved in H2O):
- For a vial with 2 mg of carboxyatractyloside: dissolve in 1065 ยตL H2O.
- Divide into 100 ยตL aliquots.
- Store at -20 ยฐC.
- O2k manual titrations ยป MiPNet09.12 O2k-Titrations
- Titration volume: 1 ยตL using a 10 ยตL Hamilton syringe (2 mL O2k-chamber).
- Final concentration: 1 ยตM.
- Comment: Cat is considered to be much more specific than Atractyloside (Atr), and the effective concentration required is much much lower (ยตM). With Atr, a very high concentration (mM) is required for full inhibition. Cat appears to be little permeable to intact cells and can thus only be applied in permeabilized cells and tissues or in isolated mitochondria (Ruas 2016 PLOS_ONE).
ยป Talk:Carboxyatractyloside