Difference between revisions of "Carboxyatractyloside"
From Bioblast
| Line 21: | Line 21: | ||
::::# Weigh 4.7 mg carboxyatractyloside. | ::::# Weigh 4.7 mg carboxyatractyloside. | ||
::::# Dissolve in 1 | ::::# Dissolve in 1 mL H<sub>2</sub>O. | ||
::::# Divide into 0.2 | ::::# Divide into 0.2 mL aliquots. | ||
::::# Store at -20 °C. | ::::# Store at -20 °C. | ||
::: '''O2k manual titrations''' » [[MiPNet09.12 O2k-Titrations]] | ::: '''O2k manual titrations''' » [[MiPNet09.12 O2k-Titrations]] | ||
::::* Titration volume: 2 | ::::* Titration volume: 2 µL using a 10 µL Hamilton syringe (2 mL O2k-chamber). | ||
::::* Final concenteration: 5 µM. | ::::* Final concenteration: 5 µM. | ||
| Line 33: | Line 33: | ||
:::: '''Preparation of 2 mM stock solution''' (dissolved in H<sub>2</sub>O): | :::: '''Preparation of 2 mM stock solution''' (dissolved in H<sub>2</sub>O): | ||
::::# For a vial with 2 mg of carboxyatractyloside: dissolve in 1065 | ::::# For a vial with 2 mg of carboxyatractyloside: dissolve in 1065 µL H<sub>2</sub>O. | ||
::::# Divide into 100 µL aliquots. | ::::# Divide into 100 µL aliquots. | ||
::::# Store at -20 °C. | ::::# Store at -20 °C. | ||
| Line 39: | Line 39: | ||
::: '''O2k manual titrations''' » [[MiPNet09.12 O2k-Titrations]] | ::: '''O2k manual titrations''' » [[MiPNet09.12 O2k-Titrations]] | ||
::::* Titration volume: 1 | ::::* Titration volume: 1 µL using a 10 µL Hamilton syringe (2 mL O2k-chamber). | ||
::::* Final concenteration: 1 µM. | ::::* Final concenteration: 1 µM. | ||
Revision as of 09:51, 3 September 2019
- high-resolution terminology - matching measurements at high-resolution
Carboxyatractyloside
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 °C; 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 °C, 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 °C.
- O2k manual titrations » MiPNet09.12 O2k-Titrations
- Titration volume: 2 µL using a 10 µL Hamilton syringe (2 mL O2k-chamber).
- Final concenteration: 5 µM.
- Preparation of 2 mM stock solution (dissolved in H2O):
- For a vial with 2 mg of carboxyatractyloside: dissolve in 1065 µL H2O.
- Divide into 100 µL aliquots.
- Store at -20 °C.
- O2k manual titrations » MiPNet09.12 O2k-Titrations
- Titration volume: 1 µL using a 10 µL Hamilton syringe (2 mL O2k-chamber).
- Final concenteration: 1 µM.
- Comment: Cat is considered to be much more specific than Atractyloside (Atr), and the effective concentration required is much much lower (µM). 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).
» Talk:Carboxyatractyloside
