Oxaloacetate Colorimetric Assay Kit

$ 442⁹⁵

Oxaloacetate Colorimetric Assay Kit

SKU #	E-BC-K901-M
Detection Instrument	Microplate reader (402-422 nm, optimum wavelength: 412 nm)
Detection method	Colorimetric method

Product Details

Properties

Sample type	Animal tissue
Sensitivity	0.017 mmol/L
Detection range	0.017-1 mmol/L
Detection method	Colorimetric method
Assay type	Quantitative
Assay time	15 min
Precision	Average inter-assay CV: 2.300% \| Average intra-assay CV: 2.100%
Other instruments required	Normal saline (0.9% NaCl)
Other reagents required	Incubator, Ultrafiltration tube (50kD)
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 brain tissue homogenate	1
10% Rat lung tissue homogenate	1
10% Mouse liver tissue homogenate	1
10% Mouse kidney tissue homogenate	1
10% Mouse leg muscle tissue homogenate	1
10% Pig heart tissue homogenate	1
10% Human urine 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

Oxaloacetic acid (OAA), one of the intermediates in the tricarboxylic acid cycle is an important substance in carbon and nitrogen metabolism. Oxaloacetate reacts with substrate acetyl-coA under the action of enzyme. The substance produced can react with DTNB, which has the maximum absorption peak at 412 nm. The content of oxaloacetate can be determined by measuring the absorbance value.

Kit Components & Storage

Item	Component	Size 1(48 T)	Size 2(96 T)	Storage
Reagent 1	Buffer Solution	10 mL × 1 vial	20 mL × 1 vial	-20℃, 12 months
Reagent 2	Substrate	1 mL × 1 vial	2 mL × 1 vial	-20℃, 12 months, shading light
Reagent 3	Enzyme Reagent	Liquid × 1 vial	Liquid × 2 vials	-20℃, 12 months, shading light
Reagent 4	Chromogenic Agent	1.2 mL × 1 vial	2.4 mL × 1 vial	-20℃, 12 months, shading light
Reagent 5	Standard	Power × 1 vial	Power × 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 rat liver 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)	0.06	0.24	0.75
%CV	2.5	2.2	1.6

Inter-assay Precision

Three rat liver 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)	0.06	0.24	0.75
%CV	2.4	2.2	2.3

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	0.45	0.7
Observed Conc. (mmol/L)	0.2	0.4	0.7
Recovery rate(%)	101	99	103

Sensitivity

The analytical sensitivity of the assay is 0.017 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	0.8	1.0
Average OD	0.278	0.333	0.405	0.470	0.610	0.680	0.813	0.945
Absoluted OD	0	0.055	0.127	0.192	0.332	0.402	0.535	0.667