Difference between revisions of "SF6847"

Latest revision as of 11:57, 22 June 2021

high-resolution terminology - matching measurements at high-resolution

SF6847

Description

SF6847 (C₁₈H₂₂N_2O), also known as tyrphostin A9 or malonoben, is a protonophore and a very potent uncoupler of oxidative phosphorylation, being used in the nM range. Like all uncouplers, SF6847 concentrations must be titrated carefully to evaluate the optimum concentration for maximum stimulation of mitochondrial respiration, particularly to avoid inhibition of respiration at higher concentrations.

Abbreviation: SF6847

Reference: Chinopoulos 2014 Methods Enzymol, Terada 1984 Biochim Biophys Acta

Application in HRR

SF6847: Tyrphostin A9 or Malonoben (3,5-bis(1,1-Dimethylethyl)-4-hydroxyphenylmethylenepropanedinitrile; C₁₈H₂₂N₂O), Sigma-Aldrich: T182, store at RT, CAS: 10537-47-0, M = 282.38 g·mol^-1

Hazard statements: H301, H311; toxic if swallowed, toxic in contact with skin

Preparation of 25 mM stock solution (dissolved in ethanol abs.):

Weigh 5 mg in a dark glass vial, dissolve in 708.3 µL ethanol.
Store in dark glass vials at -20 °C.

Preparation of 50 µM stock solution (dissolved in ethanol abs.) for 2-mL O2k-chamber:

Mix 998 µL ethanol abs. and 2 µL of the 25 mM stock solution.
Divide into 0.1 ml aliquotes, use dark glass vials.
Store at -20 °C.

» O2k manual titrations MiPNet09.12 O2k-Titrations

Titration volume (2-mL O2k-chamber): 1 µL steps of 50 µM stock solution using a 10 µL Hamilton syringe.
Final concentration: 25 nM steps.

Preparation of 12.5 µM stock solution (dissolved in ethanol abs.) for 0.5-mL O2k-chamber:

Add 300 µL ethanol abs. and 100 µL of the 50 µM stock solution.
Divide into 0.1 ml aliquots in dark glass vials. Store at -20 °C.

» O2k manual titrations MiPNet09.12 O2k-Titrations

Titration volume (0.5-mL O2k-chamber): 1 µL of 12.5 µM stock solution using a 10 µL Hamilton syringe.
Final concentration: 25 nM steps.

Caution: Chemicals stored in the fridge or freezer should be allowed to reach room temperature before opening.

Bioblast links: Uncoupling - >>>>>>> - Click on [Expand] or [Collapse] - >>>>>>>

Specific

» Artefacts by single dose uncoupling

» ATP synthase

» CCCP

» Coupling-control protocol

» DNP

» Dyscoupled respiration

» FCCP

» Is respiration uncoupled - noncoupled - dyscoupled?

» Noncoupled respiration: Discussion

» Uncoupler

» Uncoupled respiration - see » Noncoupled respiration

» Uncoupling proteins

» Uncoupling protein 1

» Uncoupler titrations - Optimum uncoupler concentration

Respiratory states and control ratios

» Biochemical coupling efficiency

» Coupling-control state

» Electron-transfer-pathway state

» Electron-transfer pathway

ET capacity

» E-L coupling efficiency

» Flux control efficiency

» Flux control ratio

» LEAK-control ratio

» LEAK respiration

» Noncoupled respiration

» OXPHOS

» OXPHOS capacity; » State 3

» OXPHOS-control ratio, P/E ratio

» Respiratory acceptor control ratio

» ROUTINE-control ratio

» ROUTINE respiration

» ROUTINE state

» State 3u

» State 4

» Uncoupling-control ratio UCR

Gnaiger E et al ― MitoEAGLE Task Group (2020) Mitochondrial physiology. Bioenerg Commun 2020.1. https://doi.org/10.26124/bec:2020-0001.v1
Gnaiger E (2020) Mitochondrial pathways and respiratory control. An introduction to OXPHOS analysis. 5th ed. Bioenerg Commun 2020.2. https://doi.org/10.26124/bec:2020-0002

General (alphabetical order)

» Adenine nucleotide translocase

» Adenylates

» Electron transfer pathway

» Mitochondrial preparations

» mt-membrane potential

» Oxygen flux

» Phosphorylation system

» Proton leak

» Proton slip

» TIP2k

Other keyword lists

» Template:Keywords: Force and membrane potential

MitoPedia topics: Uncoupler

@@ Line 5: / Line 5: @@
 }}
 == Application in [[HRR]] ==
-=== Preparation ===
+{{Chemical_description
-:::: '''SF 6847'''/tyrphostin A9/malonoben (C<sub>18</sub>H<sub>22</sub>N<sub>2O</sub>, [[3,5-bis(1,1-Dimethylethyl)-4-hydroxyphenyl]methylene]propanedinitrile; Sigma T182, 5 mg: M = 282.38 g·mol<sup>-1</sup>. Alternative source: Enzo, BML-EI215.
+|abbr=SF6847
+|trivial name=Tyrphostin A9 or Malonoben
+|complete name=3,5-bis(1,1-Dimethylethyl)-4-hydroxyphenylmethylenepropanedinitrile
+|chem formula=C<sub>18</sub>H<sub>22</sub>N<sub>2</sub>O
+|molar mass=282.38
+|vendor=Sigma-Aldrich
+|product number=T182
+|store at=RT
+|sensitivity=
+|cas=10537-47-0
+|h statements=H301, H311
+|h info=toxic if swallowed, toxic in contact with skin
+}}<!--:::: '''SF 6847'''/tyrphostin A9/malonoben (C<sub>18</sub>H<sub>22</sub>N<sub>2</sub>O, [[3,5-bis(1,1-Dimethylethyl)-4-hydroxyphenyl]methylene]propanedinitrile; Sigma T182, 5 mg: M = 282.38 g·mol<sup>-1</sup>. Alternative source: Enzo, BML-EI215.
+:::: <span style="color:#8B008B"> '''Caution:''' Toxic!</span>-->
-:::: <span style="color:#8B008B"> '''Caution:''' Toxic!</span>
-::: '''Preparation of 50 mM stock solution''' (dissolved in 99.9% ethanol):
+::: '''Preparation of 25 mM stock solution''' (dissolved in ethanol abs.):
-::::# With 5 mg in a dark glass vial: dissolve with 354.1 µL ethanol.
+::::# Weigh 5 mg in a dark glass vial, dissolve in 708.3 µL ethanol.
-::::# Store in glass vials at -20 °C.
+::::# Store in dark glass vials at -20 °C.
-::: '''Preparation of 500 µM stock solution''' (dissolved in 99.9% ethanol):
-::::# Add 990 µL ethanol and 10 µL of the 50 mM stock solution to a dark glass vial.
-::::# Store in glass vials at -20 °C.
-::: '''Preparation of 50 µM stock solution''' (dissolved in 99.9% ethanol):
+::: '''Preparation of 50 µM stock solution''' (dissolved in ethanol abs.) for '''2-mL O2k-chamber''':
-::::# Add 900 µL ethanol and 100 µL of the 500 µM stock solution to a dark glass vial.
+::::# Mix 998 µL ethanol abs. and 2 µL of the 25 mM stock solution.
-::::# Divide into 0.1 ml aliquotes, in dark glass vials.
+::::# Divide into 0.1 ml aliquotes, use dark glass vials.
 ::::# Store at -20 °C.
-::: '''O2k manual titrations'''  [[MiPNet09.12 O2k-Titrations]]
+::: » '''O2k manual titrations''' [[MiPNet09.12 O2k-Titrations]]
-::::* Titration volume: 1 µl steps 50 µM stock solution using a 10 µL syringe (2 ml O2k-Chamber).
+::::* Titration volume ('''2-mL O2k-chamber'''): 1 µL steps of 50 µM stock solution using a 10 µL Hamilton syringe.
 ::::* Final concentration: 25 nM steps.
+::: '''Preparation of 12.5 µM stock solution''' (dissolved in ethanol abs.)  for '''0.5-mL O2k-chamber''':
+::::# Add 300 µL ethanol abs. and 100 µL of the 50 µM stock solution.
+::::# Divide into 0.1 ml aliquots in dark glass vials. Store at -20 °C.
+::: » '''O2k manual titrations''' [[MiPNet09.12 O2k-Titrations]]
+::::* Titration volume ('''0.5-mL O2k-chamber'''): 1 µL of 12.5 µM stock solution using a 10 µL Hamilton syringe.
+::::* Final concentration: 25 nM steps.
 :::: '''Caution:''' Chemicals stored in the fridge or freezer should be allowed to reach room temperature before opening.