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ELISA Kit for Clusterin (CLU) Homo sapiens (Human) Sandwich ELISA

CLU; TRPM2; KUB1; CLI; APOJ; SGP2; AAG4; APO-J; SP-40; Apolipoprotein J; Complement Lysis Inhibitor; 40,Sulfated Glycoprotein 2; Testosterone-Repressed Prostate Message 2

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  • ELISA Kit for Clusterin (CLU) Packages (Simulation)
  • ELISA Kit for Clusterin (CLU) Packages (Simulation)
  • ELISA Kit for Clusterin (CLU) Results demonstration
  • SEB180Hu.jpg Typical Standard Curve
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

Recovery

Matrices listed below were spiked with certain level of recombinant Clusterin (CLU) and the recovery rates were calculated by comparing the measured value to the expected amount of Clusterin (CLU) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 97-105 101
EDTA plasma(n=5) 78-105 93
heparin plasma(n=5) 78-95 86

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Clusterin (CLU) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Clusterin (CLU) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Clusterin (CLU) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

Sample 1:2 1:4 1:8 1:16
serum(n=5) 87-96% 81-101% 92-99% 91-102%
EDTA plasma(n=5) 94-105% 87-95% 85-93% 80-88%
heparin plasma(n=5) 92-101% 89-103% 92-101% 86-103%

Stability

The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.

Reagents and materials provided

Reagents Quantity Reagents Quantity
Pre-coated, ready to use 96-well strip plate 1 Plate sealer for 96 wells 4
Standard 2 Standard Diluent 1×20mL
Detection Reagent A 1×120µL Assay Diluent A 1×12mL
Detection Reagent B 1×120µL Assay Diluent B 1×12mL
TMB Substrate 1×9mL Stop Solution 1×6mL
Wash Buffer (30 × concentrate) 1×20mL Instruction manual 1

Assay procedure summary

1. Prepare all reagents, samples and standards;
2. Add 100µL standard or sample to each well. Incubate 1 hours at 37°C;
3. Aspirate and add 100µL prepared Detection Reagent A. Incubate 1 hour at 37°C;
4. Aspirate and wash 3 times;
5. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
6. Aspirate and wash 5 times;
7. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
8. Add 50µL Stop Solution. Read at 450nm immediately.

ELISA Kit for Clusterin (CLU)

Test principle

The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Clusterin (CLU). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Clusterin (CLU). Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Clusterin (CLU), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of Clusterin (CLU) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Citations

  • Deficiency of Clusterin Exacerbates High-Fat Diet-Induced Insulin Resistance in Male Mice Pubmed: 24684302
  • Time-Dependent Decrease of Clusterin as a Potential Cerebrospinal Fluid Biomarker for Drug-Resistant Epilepsy.Pubmed: 24488574
  • Serum Amyloid A and Clusterin as Potential Predictive Biomarkers for Severe Hand, Foot and Mouth Disease by 2D-DIGE Proteomics AnalysisPubmed:25268271
  • Proteomic analysis of human plasma and peripheral blood mononuclear cells in Systemic Lupus Erythematosus patients.pubmed:28390925
  • Proteomics Analysis of Blood Serums from Alzheimer's Disease Patients Using iTRAQ LabelingTechnology.pubmed:27911324
  • Regulatory effects of simvastatin and apoJ on APP processing and amyloid-β clearance in blood-brain barrier endothelial cellspubmed:28941799
  • Clusterin, a New Cerebrospinal Fluid Biomarker in Severe Subarachnoid Hemorrhage: A Pilot Studypubmed:28803177
  • Screening for immune-potentiating antigens from hepatocellular carcinoma patients after radiofrequency ablation by serum proteomic analysisPubmed:29386009
  • High serum clusterin levels are correlated with premature coronary artery disease in a Chinese populationPubmed: 30659732
  • Discovery and validation of FBLN1 and ANT3 as potential biomarkers for early detection of cervical cancer33602229

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