产品资料

ELISAGenie/Human Bcl-2 (B-cell Leukemia/Lymphoma 2) CLIA Kit (HUES00109)/B-cell Leukemia/Lymphoma 2) CLIA Kit (HUES00109/HUES00109

Description
Additional Information

Description

Human Bcl-2 (B-cell Leukemia/Lymphoma 2) CLIA Kit

This kit uses Sandwich-CLIA as the method. The micro CLIA plate provided in this kit has been pre-coated with an antibody specific to Human Bcl-2 . Standards or samples are added to the appropriate micro CLIA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Human Bcl-2 and Avidin-Horseradish Peroxidase (HRP) conjugate are added to each micro plate well successively and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Human Bcl-2, biotinylated detection antibody and Avidin-HRP conjugate will appear fluorescence. The Relative light unit (RLU) value is measured spectrophotometrically by the Chemiluminescence immunoassay analyzer. The RLU value is positively associated with the concentration of Human Bcl-2. The concentration of Human Bcl-2 in the samples can be calculated by comparing the RLU of the samples to the standard curve.

Assay type	Sandwich
Format	96T
Assay time	4.5h
Reactivity	Human
Detection method	Chemiluminescence
Detection range	62.50-4000 pg/mL
Sensitivity	37.50 pg/mL
Sample volume	100uL
Sample type	Serum, plasma and other biological fluids
Repeatability	CV < 15%

Specificity

This kit recognizes Human Bcl-2in samples. No significant cross-reactivity or interference between Human Bcl-2and analogues was observed.

Typical Data

As the RLU values of the standard curve may vary according to the conditions of the actual assay performance (e.g. operator, pipetting technique, washing technique or temperature effects), the operator should establish a standard curve for each test. Typical standard curve and data is provided below for reference only.

Concentration(pg/mL)	RLU	Average	Corrected
4000	5753460628	59081	59046
2000	2528930429	27859	27824
1000	1428612682	13484	13449
500	65146696	6605	6570
250	33643122	3243	3208
125	16931469	1581	1546
62.50	725785	755	720
0	3535	35	--

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human Bcl-2were tested 20 times on one plate, respectively.

Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human Bcl-2were tested on 3 different plates, 20 replicates in each plate.

	Intra-assay Precision			Inter-assay Precision
Sample	1	2	3	1	2	3
n	20	20	20	20	20	20
Mean (pg/mL)	214.89	456.63	1932.39	213.61	468.80	1808.51
Standard deviation	20.93	44.89	182.61	20.74	50.86	184.65
C V (%)	9.74	9.83	9.45	9.71	10.85	10.21

Recovery

The recovery of Human Bcl-2spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.

Sample Type	Range (%)	Average Recovery (%)
Serum (n=5)	86-98	92
EDTA plasma (n=5)	96-111	104
Cell culture media (n=5)	93-104	98

Linearity

Samples were spiked with high concentrations of Human Bcl-2and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.

		Serum (n=5)	EDTA plasma (n=5)	Cell culture media (n=5)
1:2	Range (%)	91-106	91-109	99-112
1:2	Average (%)	97	99	106
1:4	Range (%)	91-107	89-99	98-112
1:4	Average (%)	98	94	104
1:8	Range (%)	89-106	95-109	96-111
1:8	Average (%)	97	101	103
1:16	Range (%)	97-114	89-104	85-96
1:16	Average (%)	104	95	91

Kit Components & Storage

An unopened kit can be stored at 4"C for 1 month. If the kit is not used within 1 month, store the items separately according to the following conditions once the kit is received.

Item	Specifications	Storage
Micro CLIA Plate(Dismountable)	8 wells X12 strips	-20"C, 6 months
Reference Standard	2 vials
Concentrated Biotinylated Detection Ab (100X)	1 vial, 120 uL
Concentrated HRP Conjugate (100X)	1 vial, 120 uL	-20"C(shading light), 6 months
Reference Standard & Sample Diluent	1 vial, 20 mL	4"C, 6 months
Biotinylated Detection Ab Diluent	1 vial, 14 mL
HRP Conjugate Diluent	1 vial, 14 mL
Concentrated Wash Buffer (25X)	1 vial, 30 mL
Substrate Reagent A	1 vial, 5 mL	4"C (shading light)
Substrate Reagent B	1 vial, 5 mL	4"C (shading light)
Plate Sealer	5 pieces
Product Description	1 copy
Certificate of Analysis	1 copy

Human Bcl-2 (B-cell Leukemia/Lymphoma 2) CLIA Kit(HUES00109) Assay procedure

1. Set standard, test sample and control (zero) wells on the pre-coated plate and record theirpositions. It is recommended to measure each standard and sample in duplicate. Note: addall solutions to the bottom of the plate wells while avoiding contact with the well walls. Ensuresolutions do not foam when adding to the wells.

2. Aliquot 100µl of standard solutions into the standard wells.

3. Add 100µl of Sample / Standard dilution buffer into the control (zero) well.

4. Add 100µl of properly diluted sample (serum, plasma, tissue homogenates and otherbiological fluids.) into test sample wells.

5. Cover the plate with the sealer provided in the kit and incubate for 90 min at 37°C.

6. Aspirate the liquid from each well, do not wash. Immediately add 100µL of BiotinylatedDetection Ab working solution to each well. Cover the plate with a plate seal and gently mix.Incubate for 1 hour at 37°C.

7. Aspirate or decant the solution from the plate and add 350µL of wash buffer to each welland incubate for 1-2 minutes at room temperature. Aspirate the solution from each well andclap the plate on absorbent filter paper to dry. Repeat this process 3 times. Note: a microplatewasher can be used in this step and other wash steps.

8. Add 100µL of HRP Conjugate working solution to each well. Cover with a plate seal andincubate for 30 min at 37°C.

9. Aspirate or decant the solution from each well. Repeat the wash process for five times asconducted in step 7.

10. Add 100µL of Substrate mixture solution to each well. Cover with a new plate seal andincubate for no more than 5 min at 37°C. Protect the plate from light.

11. Determine the RLU value of each well immediately.

View AllClose

Additional Information

Reactivity:	Human
ELISA Type:	CLIA