Multiplex Assay Kit for Thrombospondin 1 (THBS1) ,etc. by CBA (Cytometric Bead Array)

TSP1; Thrombospondin-1p180

Specificity

This assay has high sensitivity and excellent specificity for detection of Thrombospondin 1 (THBS1) ,etc. by CBA (Cytometric Bead Array).
No significant cross-reactivity or interference between Thrombospondin 1 (THBS1) ,etc. by CBA (Cytometric Bead Array) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Thrombospondin 1 (THBS1) ,etc. by CBA (Cytometric Bead Array) and the recovery rates were calculated by comparing the measured value to the expected amount of Thrombospondin 1 (THBS1) ,etc. by CBA (Cytometric Bead Array) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 78-88 81
EDTA plasma(n=5) 88-104 99
heparin plasma(n=5) 91-105 101

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Thrombospondin 1 (THBS1) ,etc. by CBA (Cytometric Bead Array) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Thrombospondin 1 (THBS1) ,etc. by CBA (Cytometric Bead Array) 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 Thrombospondin 1 (THBS1) ,etc. by CBA (Cytometric Bead Array) 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) 86-93% 84-92% 78-90% 90-104%
EDTA plasma(n=5) 81-101% 93-101% 93-101% 93-103%
heparin plasma(n=5) 93-102% 81-96% 79-98% 83-102%

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

Assay procedure summary

1. Add 200μL analysis buffer solution to each well of the plate for pre-wetting;
2. Preparation of standards, reagents and samples before the experiment;
3. Add 100μL standard or sample to each well, add 10μL magnetic beads, and incubate 120 minutes protect from light at 25°C on shaker;
4. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent A. Incubate 60 minutes protect from light at 25°C on shaker;
5. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent B, and incubate 30 minutes protect from light at 25°C on shaker;
6. Wash plate once on magnetic frame;
7. Add 100μL sheath fluid, swirl for 10 minutes, read on the machine.

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Magazine Citations
Journal of Physiology and Biochemistry Glucose and insulin modify thrombospondin 1 expression and secretion in primary adipocytes from diet-induced obese rats SpringerLink: 1v552w21p7423764
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PloS one Anthocyanin-Rich Purple Corn Extract Inhibit Diabetes-Associated Glomerular Angiogenesis Pubmed: 24278186
BMC Complementary and Alternative Medicine Identification of plasma protein markers common to patients with malignant tumour and Abnormal Savda in Uighur medicine: a prospective clinical study Pubmed:25652121
Oncotarget Novel circulating peptide biomarkers for esophageal squamous cell carcinoma revealed by a magnetic bead-based MALDI-TOFMS assay pubmed:26993605
Osteoarthritis and Cartilage Orosomucoid 2 serves for predicting but not for monitoring drug response in patients with knee osteoarthritis treated with chondroitin sulfate/glucosamine … 10.1016:j.joca.2018.02.412
GEBURTSHILFE UND FRAUENHEILKUNDE Prediction of Spontaneous Preterm Birth in At-risk Women Using Thrombospondin 1 from Cervicovaginal Fluid: A Prospective Observational Study 34531612
NPJ Regen Med Mesenchymal stromal cells mitigate liver damage after extended resection in the pig by modulating thrombospondin-1/TGF-β 34862411
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