Adenosine Diphosphate Glucose Prophosphorylase (AGP) Activity Assay Kit

$ 387⁹⁵

Adenosine Diphosphate Glucose Prophosphorylase (AGP) Activity Assay Kit

SKU #	E-BC-K856-M
Detection Instrument	Microplate reader (440-460 nm, optimum wavelength: 450 nm)
Detection method:	Colorimetric method

Product Details

Properties

Synonyms	AGP
Sample type	Plant tissue
Sensitivity	0.5 U/L
Detection range	0.5-81.59 U/L
Detection method	Colorimetric method
Assay type	Enzyme Activity
Precision	Average inter-assay CV: 5.5-10% \| Average intra-assay CV: 3.0-4.2%
Other instruments required	Vortex mixer, Incubator
Other reagents required	Normal saline (0.9% NaCl)
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% Corn tissue homogenate	1
10% Epipremnum aureum tissue homogenate	1
10% Asparagus tissue homogenate	1
10% Barley tissue homogenate	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

Adenosine diphosphate glucose pyrophosphorylase (AGP) catalyzes the substrate to produce an intermediate product, which produce chromogenic substances after enzyme catalysis. The substance has a maximum absorption peak at around 450 nm, and the AGP activity can be calculated by the OD value at 450 nm.

Kit Components & Storage

Item	Component	Size (96 T)	Storage
Reagent 1	Buffer Solution	20 mL × 1 vial	-20℃, 12 months
Reagent 2	Enzymatic Reagent	1.56 mL × 1 vial	-20℃, 12 months
Reagent 3	Substrate A	Powder ×1 vial	-20℃, 12 months
Reagent 4	Substrate B	2.5 mL × 1 vial	-20℃, 12 months
Reagent 5	Accelerant A	2.5 mL × 1 vial	-20℃, 12 months
Reagent 6	Chromogenic Agent	2.5 mL × 1 vial	-20℃, 12 months, shading light
Reagent 7	Accelerant B	2.5 mL × 1 vial	-20℃, 12 months
Reagent 8	0.5 mmol/L Standard Solution	3.3 mL ×1 vial	-20℃, 12 months
	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 barley tissue 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)	4.80	26.70	66.50
%CV	4.2	3.3	3.0

Inter-assay Precision

Three barley tissue 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)	4.80	26.70	66.50
%CV	5.5	10.0	7.6

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 99%.

	Standard 1	Standard 2	Standard 3
Expected Conc. (mmol/L)	0.13	0.25	0.38
Observed Conc. (mmol/L)	0.1	0.2	0.4
Recovery rate(%)	102	98	97

Sensitivity

The analytical sensitivity of the assay is 0.5 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 (mmol/L)	0	0.1	0.15	0.2	0.3	0.35	0.4	0.5
OD Value	0.083	0.212	0.284	0.353	0.494	0.562	0.629	0.783
OD Value	0.078	0.218	0.285	0.355	0.494	0.545	0.627	0.778
Average OD	0.081	0.215	0.285	0.354	0.494	0.554	0.628	0.781
Absoluted OD	0.000	0.135	0.204	0.274	0.414	0.473	0.548	0.700