Glutamic Acid (Glu) Colorimetric Assay Kit

$ 272⁹⁵

Glutamic Acid (Glu) Colorimetric Assay Kit

SKU #	E-BC-K903-M
Detection Instrument	Microplate reader (440-460 nm)

Product Details

Properties

Synonyms	Glu
Sample type	Serum (plasma), urine, animal and plant tissue and cells
Sensitivity	0.006 mmol/L
Detection range	0.006–0.8 mmol/L
Detection method	Colorimetric method
Assay type	Quantitative
Assay time	35 min
Precision	Average inter-assay CV: 4.400% \| Average intra-assay CV: 0.800%
Storage	-20℃
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% Rat liver tissue homogenate	1
10% Rat lung tissue homogenate	1
10% Rat brain tissue homogenate	1-2
10% Mouse liver tissue homogenate	1
10% Rat spleen tissue homogenate	1
10% Rat pancreatic tissue homogenate	1
10% Bovine liver tissue homogenate	1
10% Porcine heart tissue homogenate	1
Human serum	3-5
Human urine	1

Note: The diluent is normal saline (0.9% NaCl). For the dilution of other sample types, please do pretest to confirm the dilution factor.

Detection Principle

Glutamic acid can reduce NAD+ to NADH. NADH, under the action of hydrogen transmitter, reduces WST-8 to form yellow product, which has a characteristic absorption peak at 450 nm. Glutamic acid content can be calculated by measuring the OD value at 450 nm.

Kit Components & Storage

Item	Component	Size 1(48 T)	Size 2(96 T)	Storage
Reagent 1	Buffer Solution	8 mL × 1 vial	16 mL × 1 vial	-20℃, 12 months, shading light
Reagent 2	Enzyme Reagent	Powder × 1 vial	Powder × 2 vials	-20℃, 12 months, shading light
Reagent 3	Substrate	Powder × 1 vial	Powder × 2 vials	-20℃, 12 months, shading light
Reagent 4	Chromogenic Agent	1.2 mL × 1 vial	1.2 mL × 2 vials	-20℃, 12 months, shading light
Reagent 5	50 mmol/L Standard	1 mL × 1 vial	1 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 (mmol/L)	0.04	0.15	0.45
%CV	1.0	0.8	0.6

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 (mmol/L)	0.04	0.15	0.45
%CV	4.3	4.1	4.8

Recovery

Take three samples of high concentration, middle concentration and low concentration to test the samples of each concentration for 6 times parallelly to get the average recovery rate of 98%.

	Standard 1	Standard 2	Standard 3
Expected Conc. (mmol/L)	0.15	0.4	0.65
Observed Conc. (mmol/L)	0.1	0.4	0.7
Recovery rate(%)	95	99	100

Sensitivity

The analytical sensitivity of the assay is 0.006 mmol/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 (mmol/L)	0	0.1	0.2	0.3	0.5	0.6	0.7	0.8
Average OD	0.053	0.203	0.381	0.558	0.904	1.081	1.213	1.410
Absoluted OD	0.000	0.150	0.329	0.505	0.852	1.028	1.160	1.357