DTPA: Difference between revisions

Revision as of 15:55, 4 April 2024

high-resolution terminology - matching measurements at high-resolution

DTPA

Description

[[Description::DTPA (Diethylenetriamine-N,N,N',N,N-pentaacetic acid, pentetic acid,(Carboxymethyl)imino]bis(ethylenenitrilo)-tetra-acetic acid) is a polyaminopolycarboxylic acid (like EDTA) chelator of metal cations. DTPA wraps around a metal ion by forming up to eight bounds, because each COO- group and and N-center serves a center for chelation. With transition metals the number of bounds is less than eight. The compound is not cell membrane permeable. In general, it chelates multivalent ions stronger than EDTA.]]

Abbreviation: DTPA

MitoPedia methods: Fluorometry

Application in HRR

Fluorometry:

In Amplex UltraRed assay systems with HRP, there is a spontaneous increase in the UltroxRed fluorescence signal over the experimental time without any biological sample. This high background flux can be related to iron contamination. We have done a set of experiments, where we added DTPA as an iron-chelator into the O2k-chamber during AmR measurements. As a matter of fact, DTPA significantly reduced the high chemical background flux in MiR05 without affecting respiration.
Administration of DTPA into the O2k-chamber is recommended before all other chemicals because the iron chelation of the compound is time-dependent; it takes 10-15 min.
Experiments may also be run without DTPA.

DTPA (C₂H₂O₂); Dr.Ehrenstorfer GmbH (E 13095000) or Sigma Aldrich (D1133); CAS 67-43-6; The commercial powder can be stored at room temperature. The stock solution has to be stored at -20 °C; M: 393.35 g·mol^-1

Solubility: It is soluble in water (4,8 g/l at 25 °C), but not readily soluble in ethanol or apolar solvents.
Stability constant: see: [1]:

Metal	DTPA	EDTA
Ca(II)	10.74	10.96
Mg(II)	9.3	8.69
Fe(II)	16.5	14.3
Fe(III)	28.6	25.1
Mn(II)	15.6	14.04
Zn(II)	18.75	16.5

Preparation of the 5 mM stock solution of DTPA:

Weigh 1.967 mg of DTPA in a 2 mL glass vial.
Dissolve in 500 µL H₂O and add 6 µL of 5 M KOH to set the pH.
Heat up the solution up to 33 °C.
Add 2 µL 5 M KOH to adjust the pH ~ 8-9.
Add 500 µL H₂O to get 5 mM stock solution.
Divide into 50 µL portions in Eppendorf tubes.
Store at -20 °C.

»O2k manual titrations: MiPNet09.12 O2k-Titrations

Final concentration in O2k-chamber: 15 µM
Stock solution: 5 mM
Titration volume: 6 µL

@@ Line 3: / Line 3: @@
 |description='''DTPA''' (Diethylenetriamine-N,N,N',N,N-pentaacetic acid, pentetic acid,(Carboxymethyl)imino]bis(ethylenenitrilo)-tetra-acetic acid) is a polyaminopolycarboxylic acid (like EDTA) chelator of metal cations. DTPA wraps around a metal ion by forming up to eight bounds, because each COO- group and and N-center serves a center for chelation.  With transition metals the number of bounds is less than eight. The compound is not cell membrane permeable. In general, it chelates multivalent ions stronger than EDTA.
 }}
+{{MitoPedia concepts}}
+{{MitoPedia methods
+|mitopedia method=Fluorometry
+}}
+{{MitoPedia O2k and high-resolution respirometry}}
+{{MitoPedia topics}}
+[[File:DTPA structure.gif|right|200px|DTPA structure]]
 __TOC__
-[[File:DTPA structure.gif|right|200px|DTPA structure]]
+== Application in [[HRR]] ==
+:::'''Fluorometry:'''
+::::* In [[Amplex UltraRed]] assay systems with [[HRP]], there is a spontaneous increase in the UltroxRed fluorescence signal over the experimental time without any biological sample. This high background flux can be related to iron contamination. We have done a set of experiments, where we added DTPA as an iron-chelator into the O2k-chamber during AmR measurements. As a matter of fact, DTPA significantly reduced the high chemical background flux in MiR05 without affecting respiration.
+::::* Administration of DTPA into the O2k-chamber is recommended before all other chemicals because the iron chelation of the compound is time-dependent; it takes 10-15 min.
+::::* Experiments may also be run without DTPA.
+:::'''DTPA''' (C<sub>2</sub>H<sub>2</sub>O<sub>2</sub>); Dr.Ehrenstorfer GmbH (E 13095000) or Sigma Aldrich (D1133); CAS 67-43-6; The commercial powder can be stored at room temperature. The stock solution has to be stored at -20 °C; M: 393.35 g·mol<sup>-1</sup>
-::::* '''CAS:''' 67-43-6
-::::* '''Source:''' Dr.Ehrenstorfer GmbH (E 13095000) or Sigma Aldrich (D-1133).
-::::* '''M:''' 393.35 g·mol<sup>-1</sup>
-::::* '''Storage:''' The commercial powder can be stored at room temperature. The stock solution has to be stored at -20 °C.
-::::* '''Chemical formula:''' C<sub>2</sub>H<sub>2</sub>O<sub>2</sub>
 ::::* '''Solubility:''' It is soluble in water (4,8 g/l at 25 °C), but not readily soluble in ethanol or apolar solvents.
 ::::* '''Stability constant:''' see: [http://www.dojindo.com/Images/Product%20Photo/Chelate_Table_of_Stability_Constants.pdf]:
@@ Line 33: / Line 38: @@
 |-
 |}
-== Application in [[HRR]] ==
-:::'''Fluorometry:'''
-::::* In [[Amplex UltraRed]] assay systems with [[HRP]], there is a spontaneous increase in the UltoroxRed fluorescence signal over the experimental time without any biological sample. This high background flux can be related to iron contamination. We have done a set of experiments, where we added DTPA as an iron-chelator into the O2k-chamber during AmR measurements. As a matter of fact, DTPA significantly reduced the high chemical background flux in MiR05 without effecting the respiration.
-::::* Administration of DTPA into the O2k-chamber is recommended before all other chemicals, because the iron chelation of the compound is time-dependent; it takes 10-15 min.
-::::* Experiments may also be run without DTPA.
 ::: '''Preparation of the 5 mM stock solution of DTPA:'''
 ::::# Weigh 1.967 mg of DTPA in a 2 mL glass vial.
-::::# Dissolve in 500 µL ddH<sub>2</sub>O and add 6 µL of 5M KOH to set the pH.
+::::# Dissolve in 500 µL H<sub>2</sub>O and add 6 µL of 5 M KOH to set the pH.
 ::::# Heat up the solution up to 33 °C.
 ::::# Add 2 µL 5 M KOH to adjust the pH ~ 8-9.
-::::# Add 500 µL ddH<sub>2</sub>O to get 5 mM stock solution.
+::::# Add 500 µL H<sub>2</sub>O to get 5 mM stock solution.
-::::# The solution has to be prepared in glass vials, but after then it can be stored in Eppendorf tubes at -20 °C.
+::::# Divide into 50 µL portions in Eppendorf tubes.
+::::# Store at -20 °C.
 :::»'''O2k manual titrations:''' [[MiPNet09.12 O2k-Titrations]]
 ::::* Final concentration in O2k-chamber: 15 µM
 ::::* Stock solution: 5 mM
 ::::* Titration volume: 6 µL
-{{MitoPedia concepts
-|mitopedia concept=Recommended
-}}
-{{MitoPedia methods
-|mitopedia method=Fluorometry
-}}
-{{MitoPedia O2k and high-resolution respirometry}}
-{{MitoPedia topics}}