CLIA Kit for Thrombospondin 1 (THBS1)

TSP1; Thrombospondin-1p180

Specificity

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

Recovery

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

Matrix Recovery range (%) Average(%)
sodium citrate plasma(n=5) 78-91 83

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Thrombospondin 1 (THBS1) 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) 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) 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
sodium citrate plasma(n=5) 93-103% 89-101% 78-96% 80-101%

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
Substrate A 1×10mL Substrate B 1×2mL
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 100µL Substrate Solution. Incubate 10 minutes at 37°C;
8. Read RLU value immediately.

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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
Basic Research in Cardiology Secretome of apoptotic peripheral blood cells (APOSEC) attenuates microvascular obstruction in a porcine closed chest reperfused acute myocardial infarction model: role of platelet aggregation and vasodilation PubMed: 22899170
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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