Direct Bilirubin (DBIL) Colorimetric Assay Kit

Direct Bilirubin (DBIL) Colorimetric Assay Kit

SKU #	E-BC-K761-M
Detection Instrument	Microplate reader (565 nm)
Detection method	Colorimetric method

Product Details

Properties

Synonyms	DBIL
Sample type	Animal serum
Sensitivity	0.6 μmol/L
Detection range	0.6-50 μmol/L
Detection Method	Colorimetric method
Assay type	Quantitative
Assay time	35 min
Precision	Average inter-assay CV: 8.600% | Average intra-assay CV: 4%
Other instruments required	Micropipettor, Vortex mixer, Centrifuge
Storage	2-8℃
Valid period	12 months

Images

H Y Shen et al investigate the chemoprotective effect of decalactone on hepatic cancer. Direct bilirubin (DBIL) in rat liver was determined using DBIL colorimetric assay kit (E-BC-K761-M).

H Y Shen et al investigate the chemoprotective effect of decalactone on hepatic cancer. Direct bilirubin (DBIL) in rat liver was determined using DBIL colorimetric assay kit (E-BC-K761-M).

 

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 (0.7-50 μmol/L).

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

Sample type	Dilution factor
Human serum	1
Rat serum	1
Mouse serum	1
Rabbit serum	1
Chicken serum	1
Porcine serum	1

 

Note: The diluent is normal saline (0.9% NaCl).

 

Detection Principle

Bilirubin is one of the important components of bile. It is the degradation product of hemoglobin in various heme proteins under the action of a series of enzymes. It is important to the digestion and absorption of lipids and the formation of yellow distemper. Bilirubin comes in two forms: water-soluble and water-insoluble. Bilirubin has powerful antioxidant, anti-inflammatory and autoimmune properties. The concentration of bilirubin in human body is related to sex, drug intake, age and so on. Low serum bilirubin is directly related to diabetes, metabolic syndrome, cardiovascular disease and other pathological states. However, high bilirubin is indicative of hemolysis, jaundice, Gilbert syndrome, hepatitis, drug toxicity, and possible bile duct obstruction.

 

Kit Components & Storage

Item	Component	Size 1(48 T)	Size 2(96 T)	Storage
Reagent 1	Acid Agent	15 mL ×1 vial	30 mL ×1 vial	2-8℃, 12 months, shading light
Reagent 2	Diazonium Salt	5 mL × 1 vial	10 mL × 1 vial	2-8℃, 12 months
Reagent 3	Stop Solution	5 mL × 1 vial	5 mL × 1 vial	2-8℃, 12 months, shading light
Reagent 4	Standard	Powder × 2 vials	Powder × 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 (μmol/L)	2.50	23.40	45.50
%CV	4.5	4.1	3.4

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 (μmol/L)	2.50	23.40	45.50
%CV	8.2	8.8	8.8

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%.

	Sample 1	Sample 2	Sample 3
Expected Conc. (μmol/L)	12.5	26.5	42
Observed Conc. (μmol/L)	12.4	26.8	42.0
Recovery rate (%)	99	101	100

 

Sensitivity

The analytical sensitivity of the assay is 0.6 μmol/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	10	20	25	30	35	40	50
OD value of standard	0.039	0.079	0.125	0.152	0.176	0.197	0.216	0.274
0.037	0.079	0.125	0.148	0.173	0.202	0.219	0.271
Average OD	0.037	0.079	0.115	0.150	0.174	0.200	0.217	0.273
OD value of standard control	0.037	0.041	0.041	0.041	0.041	0.041	0.041	0.044
0.038	0.039	0.042	0.040	0.042	0.041	0.041	0.044
Average OD	0.038	0.040	0.041	0.041	0.041	0.041	0.041	0.044
Absoluted OD	0.000	0.039	0.074	0.109	0.133	0.159	0.177	0.229