Multiplex Assay Kit for Clusterin (CLU) ,etc. by FLIA (Flow Luminescence Immunoassay)

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

(Note: Up to 8-plex in one testing reaction)

Specificity

This assay has high sensitivity and excellent specificity for detection of Clusterin (CLU) ,etc. by FLIA (Flow Luminescence Immunoassay).
No significant cross-reactivity or interference between Clusterin (CLU) ,etc. by FLIA (Flow Luminescence Immunoassay) and analogues was observed.

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 97-104 101
EDTA plasma(n=5) 88-95 92
heparin plasma(n=5) 79-93 81
sodium citrate plasma(n=5) 87-95 90

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Clusterin (CLU) ,etc. by FLIA (Flow Luminescence Immunoassay) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Clusterin (CLU) ,etc. by FLIA (Flow Luminescence Immunoassay) 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) ,etc. by FLIA (Flow Luminescence Immunoassay) 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) 91-99% 82-101% 80-101% 89-97%
EDTA plasma(n=5) 91-102% 93-105% 81-90% 81-95%
heparin plasma(n=5) 92-99% 78-90% 93-101% 79-101%
sodium citrate plasma(n=5) 84-95% 95-103% 86-93% 88-95%

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
96-well plate 1 Plate sealer for 96 wells 4
Pre-Mixed Standard 2 Standard Diluent 1×20mL
Pre-Mixed Magnetic beads (22#:CLU) 1 Analysis buffer 1×20mL
Pre-Mixed Detection Reagent A 1×120μL Assay Diluent A 1×12mL
Detection Reagent B (PE-SA) 1×120μL Assay Diluent B 1×12mL
Sheath Fluid 1×10mL Wash Buffer (30 × concentrate) 1×20mL
Instruction manual 1

Assay procedure summary

1. Preparation of standards, reagents and samples before the experiment;
2. Add 100μL standard or sample to each well,
    add 10μL magnetic beads, and incubate 90min at 37°C on shaker;
3. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent A. Incubate 60min at 37°C on shaker;
4. Wash plate on magnetic frame for three times;
5. Add 100μL prepared Detection Reagent B, and incubate 30 min at 37°C on shaker;
6. Wash plate on magnetic frame for three times;
7. Add 100μL sheath solution, swirl for 2 minutes, read on the machine.

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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
Catalog No. Related products for research use of Homo sapiens (Human) Organism species Applications (RESEARCH USE ONLY!)
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