QuicKey Human CLU(Clusterin) ELISA Kit

SKU: E-TSEL-H0014

$ 331⁹⁵

QuicKey Human CLU(Clusterin) ELISA Kit

Detection Range	1.56-100 ng/mL
Sensitivity	0.94 ng/mL

Product Details

Properties

Assay type	Sandwich-ELISA
Format	96T
Assay time	2.5h
Detection range	1.56-100 ng/mL
Sensitivity	0.94 ng/mL
Sample type &Sample volume	serum, plasma, urine, saliva; 50μL
Specificity	This kit recognizes Human CLU in samples. No significant or interference between Human CLU and analogues was observed.
Reproducibility	Both intra-CV and inter-CV are < 10%.
Application	This ELISA kit applies to the in vitro quantitative determination of Human CLU concentrations in serum, plasma, urine, saliva.Please consult technical support for the applicability if other biological fluids need to be tested.

Test Principle

This ELISA kit uses the Sandwich-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Human CLU. Samples (or Standards) and biotinylated detection antibody specific for Human CLU are added to the micro ELISA plate wells. Human CLU would combine with the specific antibody. Then Avidin-Horseradish Peroxidase (HRP) conjugate are added successively to each micro plate well and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Human CLU, biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by the addition of stop solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 ± 2 nm. The OD value is proportional to the concentration of Human CLU. You can calculate the concentration of Human CLU in the samples by comparing the OD of the samples to the standard curve.

Kit Components and Storage

An unopened kit can be stored at 2-8℃ for six months. After test, the unused wells and reagents should be stored according to the table below.

Item	Specifications	Storage conditions after test
Micro ELISA Plate (Dismountable)	96T: 8 wells ×12 strips 48T: 8 wells ×6 strips	2-8℃, 1 month
Reference Standard	96T: 2 vials 48T: 1 vial	Discard unused reconstituted standard and dilutions
Reference Standard & Sample Diluent	1 vial, 20 mL	2-8℃
Biotinylated Detection Ab Working Solution	1 vial, 6 mL
HRP Conjugate Diluent	1 vial, 14 mL
Concentrated Wash Buffer (25×)	1 vial, 30 mL
Concentrated HRP Conjugate (100×)	96T: 1 vial, 120 μL 48T: 1 vial, 60 μL	2-8℃ (Protect from light)
Substrate Reagent	1 vial, 10 mL	2-8℃ (Protect from light)
Stop Solution	1 vial, 10 mL	2-8℃
Plate Sealer	5 pieces
Product Description	1 copy
Certificate of Analysis	1 copy

Technical Data:

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human CLU were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human CLU were 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 (ng/mL)	4.57	10.92	48.94	4.60	12.45	41.92
Standard deviation	0.24	0.51	1.99	0.24	0.62	1.95
CV (%)	5.25	4.67	4.07	5.22	4.98	4.65

Recovery

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

Sample Type	Range (%)	Average Recovery (%)
Serum(n=8)	90-101	97
EDTA plasma(n=8)	88-98	95
Urine(n=8)	83-96	91

Target Information

Background

Clusterin is a ubiquitous protein that is arousing increasing interest owing to its widespread diffusion and multifunctional role. It is composed of two 40 kD subments (NA1, NA2) encoded by a single gene, which hold together by disulphide bonds. The mature dimer has a molecular mass of 80 kD and is micro-heterogeneous in electrical charge due to glycosylation. This protein has been differently named as apolipoprotein J (ApoJ), sulfated glycoprotein 2 (SGP-2), glycoprotein III (GP- III), testosterone repressed prostate message (TRPM-2), SP-40, 40cytolysis inhibitor (CL1) and others [1].It is a component of the high-density lipoprotein (HDL) complex with putative functions in the transport of lipids, apoptosis, protection of cells from stress and, most important, in the regulation of complement activity. It has been postulated that clusterin is involved in a variety of human diseases including Alzheimer disease, brain injury following ischemia，nephrosis and myocarditis [2].
1. Wilson M R, Easterbrooksmith S B. Clusterin is a secreted mammalian chaperone. [J]. Trends in Biochemical Sciences, 2000, 25(3):95-98.
2. Mclaughlin L, Zhu G, Mistry M, et al. Apolipoprotein J/clusterin limits the severity of murine autoimmune myocarditis. [J]. Journal of Clinical Investigation, 2000, 106(9):1105-1113.

Database Links	SwissProt: P10909 Entrez Gene: 1191
Research Area	Cancer, Cardiovascular, Metabolism, Immunology, Signal transduction

Assay Procedures

	1. Add 50μL standard or sample to the wells, immediately add 50μL Biotinylated Detection Ab working solution to each well. Incubate for 90 min at 37°C
	2. Aspirate and wash the plate for 3 times
	3. Add 100μL HRP conjugate working solution. Incubate for 30 min at 37°C. Aspirate and wash the plate for 5 times
	4. Add 90μL Substrate Reagent. Incubate for 15 min at 37°C
	5. Add 50μL Stop Solution
	6. Read the plate at 450nm immediately. Calculation of the results