Quantification of Reduced Glutamine.

∼ Features ∼

  1. Easy extraction of cell Lysate 
  2. Used to determine intracellular levels of Glutamine 


 

Product Description

Glutamine, a source of α-ketoglutarate (one of the tricarboxylic acid (TCA) cycle intermediates), is an important substance used for energy production and the syntheses of nucleic acids and other amino acids. Glutaminolysis, the process by which glutamine as a substrate is converted to α-ketoglutarate, is upregulated in cancer cells. A study reports that glutaminolysis has largely contributed to scavenging of reactive oxygen species and reduction of oxidized glutathione.

The Glutamine Assay Kit-WST can quantify glutamine, a substrate of energy metabolism. Our Glutamine Assay Kit-WST is designed to quantify glutamine as a metabolite. This kit allows you to quantify glutamine present in culture medium or intracellular glutamine by WST reduction reaction. The lowest concentration of glutamine which can be quantified is 5 μmol/l. This kit can be used with 96-well microplates, which makes it possible to analyze multiple specimens.


Principle

The kit is used to detect Glutamine in cell culture medium by measuring the absorbance of WST formazan produced according to quantities of glutamine. The kit includes a “Glutamine Standard” which can be used to quantify the concentration of lactate found within samples by creating standard curve.


Simple Assay Procedure

*The glutamine concentration can be calculated by using the following equation:

glutamine (mmol/l) = (Glutaminase solution (+))-(Glutaminase solution (-))



Standard Curve

Standard curve can be prepared by using Glutamine Standard included in the kit. The concentration of Glutamine can be measured. The concentration of lactate can be evaluated by diluting the samples if the concentration is over 0.5 mmol/l.


Example of Measurements of Glutamine/Glutamate level

A549 cells were seeded on 6-well plates. Glutamine concentrations in the culture supernatant over incubation time were measured using the Glutamine Assay Kit-WST, and changes in glutamate concentration were measured using the Glutamate Assay Kit-WST. As a result, upregulated glutaminolysis led to a time-dependent decrease in glutamine concentrations and a time-dependent increase in glutamate concentrations in culture medium.


Storage Condition : 0-5oC, protect from light
Shipping Condition : ambient temperature

Q. How many samples can I measure?

A.The number of samples that can be recorded when the standard curve and sample is measured in triplicates.

100 tests
Sample 
(measured in triplicate
12 samples*
(See picture below)

A plate layout for Glutamate standard solution and sample

Prepare a test sample to be assayed. Please note that a minimum of 240 μl of sample (e.g., supernatant, lysate) is needed for each test sample

The glutamine concentration can be calculated by using the following equation:

glutamine (mmol/l) = (Glutaminase solution (+))-(Glutaminase solution (-))

Q. How stable is Working solution?

A. You can’t store the Working solution. Please prepare the Working solution prior to use. Please protect from light because the Working solution is light-sensitive.

Q. Can I quantify D-Glutamine levels?

A. No, the kit is used for the quantification of L-Glutamine levels.

Q. Can I measure samples that contain a reducing agent?

A. You can’t accurately measure Glutamine levels if your samples contain a reducing agent. WST dye interferes with the reducing agent and can cause errors in observations. If you add a testing material that has reducing properties, please prepare a few wells that only contains the medium (no cells) and testing material for background control.

Q. Our samples did not change in color, are there any reasons for this?

A. Glutamine level found within the sample may be lower than the detection limit (5 µmol/l) that can be measured using this kit.

If you dilute the sample, lactate level present in the diluted sample may be lower than 5 µmol/l.

Please lower the dilution ratio and adjust the concentration of the sample over the detection limit that was measured using this kit.

Item # Description/Size Availability Qty Break Price Quantity
G268-10
100 tests
8-14 business days 1 $550.00

*Your estimated shipping date is calculated based on production, payment method, destination, and your shipping option.
For Research Use Only Products