Glutaminase (GLS) Activity Assay Kit

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Glutaminase (GLS) Activity Assay Kit

SKU #	E-BC-K660-M
Detection Instrument	Microplate reader (440-460 nm, optimum wavelength: 450 nm)
Detection method	Colorimetric method

Product Details

Properties

Synonyms	GLS
Sample type	Serum (plasma), animal, plant tissue
Sensitivity	0.003 U/L
Detection range	0.003-18.0 U/L
Detection method	Colorimetric method
Assay type	Enzyme Activity
Assay time	80 min
Precision	Average inter-assay CV: 8.300% \| Average intra-assay CV: 5%
Other instruments required	Test tube, Incubator, Centrifuge
Other reagents required	PBS (0.01 mol/L, pH 7.4)
Storage
Valid period	12 months

Dilution of Sample

The recommended dilution factor for different samples is as follows (for reference only):

Sample type	Dilution factor
10% Mouse liver tissue homogenate	1
10% Rat brain tissue homogenate	1
10% Mouse kidney tissue homogenate	1
10% Rat liver tissue homogenate	1
10% Rat heart tissue homogenate 1	1
10% Rat kidney tissue homogenate	1
Human serum	1
Human plasma	1-3

Note: The diluent is PBS (0.01 M, pH 7.4). For the dilution of other sample types, please do pretest to confirm the dilution factor.

Detection Principle

Glutamine is decomposed to produce glutamic acid under the action of glutaminase. Glutamic acid is further transformed by glutamic acid dehydrogenase. Meanwhile, NAD+ is reduced to NADH, Which under the action of hydrogen transmitter, transfer electrons to WST-8 to produce the yellow product. The activity of glutaminase can be calculated by measuring the change of absorbance value at 450 nm.

Kit Components & Storage

Item	Component	Size (96 T)	Storage
Reagent 1	Substrate A	Powder × 2 vials	-20℃, 12 months
Reagent 2	Standard	Powder × 2 vials	-20℃, 12 months, shading light
Reagent 3	Diluent	4 mL×1 vial	-20℃, 12 months
Reagent 4	Enzyme Reagent	Powder × 2 vials	-20℃, 12 months, shading light
Reagent 5	Buffer Solution	20 mL×1 vial	-20℃, 12 months
Reagent 6	Substrate B	Powder × 2 vials	-20℃, 12 months, shading light
Reagent 7	Accelerator	Powder × 1 vial	-20℃, 12 months, shading light
Reagent 8	Accelerator	1.5 mL × 2 vials	-20℃, 12 months, shading light
	Microplate	96 wells	No requirement
	Plate Sealer	2 pieces

Note: The reagents must be stored strictly according to the preservation conditions in the above table. The reagents in different kits cannot be mixed with each other. For a small volume of reagents, please centrifuge before use, so as not to obtain sufficient amount of reagents.

Technical Data:

Parameter:

Intra-assay Precision

Three human serum samples were assayed in replicates of 20 to determine precision within an assay (CV = Coefficient of Variation).

Parameters	Sample 1	Sample 2	Sample 3
Mean (U/L)	0.26	2.80	13.50
%CV	5.4	5.2	4.7

Inter-assay Precision

Three human serum samples were assayed 20 times in duplicate by three operators to determine precision between assays.

Parameters	Sample 1	Sample 2	Sample 3
Mean (U/L)	0.26	2.80	13.50
%CV	8.2	7.9	8.8

Sensitivity

The analytical sensitivity of the assay is 0.003 U/L. This was determined by adding two standard deviations to the mean O.D. obtained when the zero standard was assayed 20 times, and calculating the corresponding concentration.

Standard Curve

As the OD value of the standard curve may vary according to the conditions of the actual assay performance (e.g. operator, pipetting technique or temperature effects), so the standard curve and data are provided as below for reference only:

Concentration (μmol/L)	0	0.1	0.15	0.2	0.3	0.4	0.45	0.5
Average OD	0.067	0.175	0.224	0.298	0.423	0.535	0.577	0.625
Absoluted OD	0.000	0.108	0.158	0.231	0.357	0.468	0.511	0.559