Catalase (CAT) Activity Assay Kit

To better serve you, we would like to discuss your specific requirement, Please Contact Us for a quote.

Catalase (CAT) Activity Assay Kit

SKU #	E-BC-K031-M
Detection Instrument	Microplate reader (400-410 nm, optimum wavelength: 405 nm)
Detection Method	Colorimetric method

Product Details

Properties

Synonyms	CAT
Sample Type	Serum, plasma, cells, cell culture supernatant, tissue
Sensitivity	1.12 U/mL
Detection Range	1.12 -150 U/mL
Detection Method	Colorimetric method
Assay type	Enzyme Activity
Assay time	40 min
Precision	Average inter-assay CV: 7.700% \| Average intra-assay CV: 3.900%
Other instruments required	Micropipettor, Vortex Mixer, Incubator
Other reagents required	Normal saline (0.9% NaCl) or PBS (0.01 M, pH 7.4)
Storage	2-8℃
Valid period	12 months

Dilution of Sample

It is recommended to take 2~3 samples with expected large difference to do pre-experiment before formal experiment and dilute the sample according to the result of the pre-experiment and the detection range (1.12 -150 U/mL).

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

Sample type	Dilution factor
Human serum	1
293T supernatant	1
10% Rat heart tissue homogenization	50-100
10% Rat liver tissue homogenization	100-200
10% Rat spleen tissue homogenization	50-100
Mouse serum	1
10% Epipremnum aureum tissue homogenization	1-2
10% Rat lung tissue homogenization	50-100
10% Rat kidney tissue homogenization	50-100
10% Rat brain tissue homogenization	20-50

Note: The diluent is normal saline (0.9% NaCl) or PBS (0.01 M, pH 7.4).

Detection Principle

The reaction that catalase (CAT) decomposes H2O2 can be quickly stopped by ammonium molybdate. The residual H2O2 reacts with ammonium molybdate to generate a yellowish complex. CAT activity can be calculated by production of the yellowish complex at 405 nm.

Kit Components & Storage

Item	Component	Size 1(48 T)	Size 2(96 T)	Storage
Reagent 1	Buffer Solution	12 mL × 1 vial	24 mL × 1 vial	2-8℃, 12 months
Reagent 2	Substrate	1.5 mL × 1 vial	1.5 mL × 2 vials	2-8℃, 12 months
Reagent 3	Chromogenic Agent	Powder ×1 vial	Powder ×1 vial	2-8℃, 12 months
Reagent 4	Clarificant	1.5 mL × 1 vial	1.5 mL × 2 vials	2-8℃, 12 months
Reagent 5	9.6 mol/L H2O2 Standard Solution	1.5 mL× 1 vial	1.5 mL × 2 vials	2-8℃, 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/mL)	3.70	26.40	78.60
%CV	4.2	3.8	3.7

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/mL)	3.70	26.40	78.60
%CV	7.4	7.9	7.8

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 100%.

	Standard 1	Standard 2	Standard 3
Expected Conc. (μmol/mL)	15	36	54
Observed Conc. (μmol/mL)	15.2	34.9	55.1
Recovery rate (%)	101	97	102

Sensitivity

The analytical sensitivity of the assay is 1.12 U/mL CAT. 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/mL)	0	10	20	40	50	60	80	100
Average OD	0.062	0.087	0.118	0.143	0.169	0.192	0.219	0.311
Absoluted OD	0	0.025	0.056	0.081	0.107	0.13	0.157	0.249