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Key Documents

P7275

Sigma-Aldrich

Pyrophosphate reagent

spectrophotometric assay

Synonym(s):

Pyrophosphate detection reagent

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About This Item

UNSPSC Code:
12352204
NACRES:
NA.54

Quality Level

form

powder

usage

 vial sufficient for ≤4 assays

storage temp.

−20°C

Application

The product has been used for the measurement of pyrophosphate content of plant tissues. It has also been used as a rapid, routine check of pyrophosphate content by monitoring NADH oxidation in Solanum tuberosum tuber extracts.

Biochem/physiol Actions

The pyrophosphate detection kit couples the production of pyrophosphate to the consumption of NADH.

Packaging

A technical bulletin is included.

Components

ComponentConcentration
Imidazole × HCl, pH 7.4 45 mM
Citrate 5 mM
EDTA 0.10 mM
Mg2+, Mn2+, Co2+ 2 mM, 0.2 mM, 0.02 mM
β-NADH 0.8 mM
F-6-P 12 mM
Bovine Serum Albumin 5 mg/mL
Sugar Stabilizer 5 mg/mL
PPi-PFK 0.5 units/mL
Aldolase 7.5 units/mL
GDH 5 units/mL
TPI 50 units/mL

Quality

A coupled enzyme system containing fructose-6-phosphate kinase (pyrophosphate-dependent), aldolase, triosephosphate isomerase, and α-glycerophosphate dehydrogenase.

Principle

Two moles of NADH are oxidized to NAD+ per mole of pyrophosphate consumed. The reaction is monitored spectrophotometrically at 340 nm.
The reagent can be used to detect 10-100 nmoles of pyrophosphate.

Preparation Note

Dissolve the contents of one vial of Pyrophosphate Reagent in 4.0 mL of water.

Storage Class Code

11 - Combustible Solids

WGK

WGK 3

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable

Personal Protective Equipment

dust mask type N95 (US), Eyeshields, Gloves

Certificates of Analysis (COA)

Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.

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Devin W Cochrane et al.
Plant science : an international journal of experimental plant biology, 265, 124-130 (2017-12-11)
Class 1 plant hemoglobins (phytoglobins) are upregulated during low-oxygen stress and participate in metabolism and cell signaling via modulation of the levels of nitric oxide (NO). We studied the effects of overexpression and knockdown of the class 1 phytoglobin gene
R Viola et al.
Plant physiology, 97(2), 638-643 (1991-10-01)
Pretreatment of discs excised from developing tubers of potato (Solanum tuberosum L.) with 10 millimolar sodium fluoride induced a transient increase in 3-phosphoglycerate content. This was followed by increases in triose-phosphate, fructose 1,6-bisphosphate and hexose-phosphate (glucose 6-phosphate + fructose 6-phosphate
Erick Strauss et al.
The Journal of biological chemistry, 277(50), 48205-48209 (2002-10-10)
Pantothenic acid (vitamin B(5)) is the natural precursor of coenzyme A (CoA), an essential cofactor in all organisms. The pantothenic acid antimetabolite N-pentylpantothenamide inhibits the growth of Escherichia coli with a minimum inhibitory concentration of 2 microm. In this study
A continuous spectrophotometric assay for argininosuccinate synthetase based on pyrophosphate formation.
W E O'Brien
Analytical biochemistry, 76(2), 423-430 (1976-12-01)
D A Smyth et al.
Plant physiology, 75(3), 862-864 (1984-07-01)
Pyrophosphate (PPi) was measured in pea (Pisum sativum L.) and corn (Zea mays L.) tissues by using an enzymic method based on PPi-dependent phosphofructokinase (PPi-PFK). Different organs of pea and corn seedlings were extracted to determine if PPi is present

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