β - Galactosidase (β –GAL) Activity Assay Kit

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β - Galactosidase (β –GAL) Activity Assay Kit

SKU #	E-BC-K631-M
Detection Instrument	Microplate reader (390-410 nm, optimum wavelength: 400 nm)
Detection method	Colorimetric method

Product Details

Properties

Synonyms	β-GAL
Sample type	Serum , plasma, cells, animal and plant tissue
Sensitivity	16.19 U/L
Detection range	16.19-1200 U/L
Detection Method	Colorimetric method
Assay type	Enzyme Activity
Assay time	70 min
Precision	Average inter-assay CV: 6% \| Average intra-assay CV: 3%
Other instruments required	Water bath, Incubator, Centrifuge, Microplate reader
Other reagents required	2 mmol/L hydrochloric acid
Storage	Reagent 2: -20℃; Others: 2-8℃
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% Rat kidney tissue homogenate	3-5
10% Rat liver tissue homogenate	1-3
10% Rat heart tissue homogenate	1
Human serum	1

Note: The diluent is buffer solution. For the dilution of other sample types, please do pretest to confirm the dilution factor.

Detection Principle

β-galactosidase can catalyze the substrate nitrophenylgalactoside pyranopyranoside to produce nitrophenol, which has a maximum absorption peak at 400 nm. The activity of β-GAL can be calculated by measuring the OD value at 400 nm.

Kit Components & Storage

Item	Component	Size (96 T)	Storage
Reagent 1	Buffer Solution	25 mL ×1 vial	2-8℃, 12 months, shading light
Reagent 2	Substrate	Powder × 1 vial	-20℃, 12 months
Reagent 3	Activator Agent	1.5 mL × 1 vial	2-8℃, 12 months shading light
Reagent 4	Chromogenic Agent	25 mL×1 vial	2-8℃, 12 months
Reagent 5	20 mmol/L Standard Solution	1 mL × 1 vial	2-8℃, 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 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)	25.60	435.00	653.00
%CV	3.2	3.0	2.8

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)	25.60	435.00	653.00
%CV	5.6	6.3	6.1

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 101%.

	Standard 1	Standard 2	Standard 3
Expected Conc. (mmol/L)	0.15	0.46	0.77
Observed Conc. (mmol/L)	0.2	0.5	0.8
Recovery rate (%)	101	99	103

Sensitivity

The analytical sensitivity of the assay is 16.19 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	0.2	0.4	0.5	0.6	0.7	0.8
Average OD	0.040	0.112	0.192	0.328	0.403	0.490	0.565	0.640
Absoluted OD	0.000	0.072	0.152	0.288	0.363	0.450	0.525	0.600