ELISA Kit for Clusterin (CLU)

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

Specificity

This assay has high sensitivity and excellent specificity for detection of Clusterin (CLU).
No significant cross-reactivity or interference between Clusterin (CLU) and analogues was observed.

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) 79-104 91
EDTA plasma(n=5) 83-99 86
heparin plasma(n=5) 89-101 95

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) 94-101% 86-101% 78-92% 78-103%
EDTA plasma(n=5) 82-92% 78-97% 80-98% 92-105%
heparin plasma(n=5) 78-99% 78-105% 98-105% 79-95%

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.

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Magazine Citations
Endocrinology Deficiency of Clusterin Exacerbates High-Fat Diet-Induced Insulin Resistance in Male Mice Pubmed: 24684302
J Mol Neurosci. Time-Dependent Decrease of Clusterin as a Potential Cerebrospinal Fluid Biomarker for Drug-Resistant Epilepsy. Pubmed: 24488574
PLOS ONE Serum Amyloid A and Clusterin as Potential Predictive Biomarkers for Severe Hand, Foot and Mouth Disease by 2D-DIGE Proteomics Analysis Pubmed:25268271
journal of immunological methods Proteomic analysis of human plasma and peripheral blood mononuclear cells in Systemic Lupus Erythematosus patients. pubmed:28390925
Journal of Alzheimer's Disease Proteomics Analysis of Blood Serums from Alzheimer's Disease Patients Using iTRAQ LabelingTechnology. pubmed:27911324
Biochimica Et Biophysica Acta Regulatory effects of simvastatin and apoJ on APP processing and amyloid-β clearance in blood-brain barrier endothelial cells pubmed:28941799
World Neurosurgery Clusterin, a New Cerebrospinal Fluid Biomarker in Severe Subarachnoid Hemorrhage: A Pilot Study pubmed:28803177
BMC Cancer Screening for immune-potentiating antigens from hepatocellular carcinoma patients after radiofrequency ablation by serum proteomic analysis Pubmed:29386009
Diabetes/metabolism research and reviews High serum clusterin levels are correlated with premature coronary artery disease in a Chinese population Pubmed: 30659732
Cancer Cell Int Discovery and validation of FBLN1 and ANT3 as potential biomarkers for early detection of cervical cancer 33602229
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