Tartrate Resistant Acid Phosphatase (TRAP) Activity Assay Kit

$ 153⁹⁵

Tartrate Resistant Acid Phosphatase (TRAP) Activity Assay Kit

SKU #	E-BC-K871-M
Detection Instrument	Microplate reader (410-415 nm, optimum wavelength: 405 nm)
Detection method:	Colorimetric method

Product Details

Properties

Synonyms	TRAP
Sample type	Serum, plasma, urine，animal tissue and cell samples
Sensitivity	9.22 U/L
Detection range	9.22-100 U/L
Detection method	Colorimetric method
Assay type	Enzyme Activity
Assay time	25 min
Precision	Average inter-assay CV: 2.300% \| Average intra-assay CV: 2%
Other instruments required	Incubator(37℃)
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
Goat plasma	1
Human urine	1
Porcine serum	1
Rat plasma	1-2
10% Mouse liver tissue homogenate	5-8
10% Mouse kidney tissue homogenate	3-5
10% Mouse lung tissue homogenate	2-5
10% Mouse brain tissue homogenate	3-5
1×10^6 HL-60 cells	1
1×10^6 293T 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

The chromogenic agent can be catalyzed by acid phosphatase to produce p-nitrophenol in acidic condition. P-nitrophenol has a maximum absorption at 405 nm. The activity of TRAP can be calculated by measuring the amount of produced p-nitrophenol. The activity of acid phosphatase being detected in the presence of tartric acid is regarded as TRAP activity.

Kit Components & Storage

Item	Component	Size (96 T)	Storage
Reagent 1	Buffer Solution	24 mL × 1 vial	-20°C, 12 months
Reagent 2	Substrate	Powder × 2 vials	-20°C, 12 months, shading light
Reagent 3	Tartaric Acid Solution	1.4 mL × 2 vials	-20℃, 12 months
Reagent 4	Standard Substance	Powder × 2 vials	-20°C, 12 months, shading light
Reagent 5	Chromogenic Agent	15 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 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)	16.80	34.50	87.00
%CV	2.3	2.0	1.7

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/L)	16.80	34.50	87.00
%CV	2.1	2.4	2.4

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)	0.25	0.52	0.84
Observed Conc. (mmol/L)	0.3	0.5	0.9
Recovery rate(%)	101	99	103

Sensitivity

The analytical sensitivity of the assay is 9.22 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	0.2	0.3	0.4	0.6	0.8	0.9	1.0
OD Value	0.079	0.25	0.332	0.417	0.601	0.767	0.852	0.953
OD Value	0.078	0.249	0.336	0.421	0.595	0.764	0.86	0.941
Average OD	0.078	0.249	0.334	0.419	0.598	0.765	0.856	0.947
Absoluted OD	0	0.171	0.256	0.340	0.519	0.687	0.777	0.868