Malic Acid Colorimetric Assay Kit

£316⁰⁰
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Malic Acid Colorimetric Assay Kit

SKU #	E-BC-K905-M
Detection Instrument	Microplate reader (445-465 nm, optimum wavelength: 450 nm)
Detection method:	Colorimetric method

Product Details

Properties

Sample type	Serum (plasma), tissue and cell
Sensitivity	0.003 mmol/L
Detection range	0.003-0.5 mmol/L
Detection method	Colorimetric method
Assay type	Quantitative
Precision	Average inter-assay CV: 4.6-5.9% \| Average intra-assay CV: 2.6-3.6%
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	3-5
10% Mouse liver tissue homogenate	3-5
10% Lemon tissue homogenate	8-12
10% Tangerine tissue homogenate	5-10
10% Potato tissue homogenate	5-12
Rat serum	1
Mouse plasma	1
3.35×10⁶ CHO cells	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

L-malic acid, also known as 2-hydroxysuccinic acid, is widely present in living organisms. It has two stereoisomers and is one of the members of the tricarboxylic acid cycle, an important metabolic cycle in living organisms. The detection principle of this kit is that the enzyme catalyzes malic acid to produce oxaloacetic acid. At the same time, under the action of electronic coupler, the chromogenic agent is reduced to produce orange yellow product, which has the maximum absorption peak at about 450 nm.

Kit Components & Storage

Item	Component	Size (96 T)	Storage
Reagent 1	Buffer Solution	20 mL × 1 vial	-20°C, 12 months
Reagent 2	Enzymatic Reagent	0.3 mL × 1 vial	-20°C, 12 months, shading light
Reagent 3	Chromogenic Agent	2.4 mL × 1 vial	-20°C, 12 months, shading light
Reagent 4	Substrate	Powder × 2 vials	-20°C, 12 months, shading light
Reagent 5	0.5 mmol/L Standard	5 mL × 1 vial	-20°C, 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)	0.06	0.10	0.30
%CV	3.6	3.4	2.6

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)	0.06	0.10	0.30
%CV	4.6	5.9	5.1

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 98%.

	Standard 1	Standard 2	Standard 3
Expected Conc. (mmol/L)	0.13	0.27	0.38
Observed Conc. (mmol/L)	0.1	0.3	0.4
Recovery rate(%)	101	96	97

Sensitivity

The analytical sensitivity of the assay is 0.003 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	0.15	0.2	0.3	0.35	0.4	0.5
Average OD	0.064	0.130	0.157	0.191	0.257	0.292	0.320	0.383
Absoluted OD	0	0.059	0.093	0.127	0.193	0.229	0.257	0.319