Multiplex Assay Kit for Sclerostin (SOST) ,etc. by FLIA (Flow Luminescence Immunoassay)

VBCH; Sclerosteosis

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

Specificity

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

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 80-101 84
EDTA plasma(n=5) 94-105 102
heparin plasma(n=5) 89-99 94

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Sclerostin (SOST) ,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 Sclerostin (SOST) ,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 Sclerostin (SOST) ,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) 89-96% 86-101% 83-102% 92-105%
EDTA plasma(n=5) 90-103% 85-105% 98-105% 94-105%
heparin plasma(n=5) 79-92% 99-105% 93-105% 79-89%

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#:SOST) 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
Life Sciences Exendin-4 Increases Bone Mineral Density in Type 2 Diabetic OLETF Rats Potentially Through the Down-Regulation of SOST/Sclerostin in Osteocytes PubMed: 23357248
Calcified Tissue International Acute effects of glucocorticoids on serum markers of osteoclasts, osteoblasts, and osteocytes Pubmed: 23247536
Arthritis Res Ther. Serum Sclerostin in Adult High-Activity Patients with Juvenile Idiopathic Arthritis Pubmed:Pmc4236454
Korean J Intern Med Early effects of tumor necrosis factor inhibition on bone homeostasis after soluble tumor necrosis factor receptor use Pubmed:Pmc4219971
Journal of Bone and Mineral Research The Anti-Osteoanabolic Function of Sclerostin is Blunted in Mice Carrying a High Bone Mass Mutation of Lrp5 Pubmed:25640331
FASEB J. DMP-1-mediated PubMed: 26481310
Pharmacology Effect of Mirtazapine on Rat Bone Tissue after Orchidectomy PubMed: 25871861
European Journal of Pharmacology The effect of levetiracetam on rat bone mineral density, bone structure and biochemical markers of bone metabolism Pubmed:29428468
EFFECTS OF DISEASE-MODIFYING ANTIRHEUMATIC DRUG TREATMENT ON PERIARTICULAR BONE REMODELING IN RHEUMATOID AND PSORIATIC … :
Journal of Clinical and Translational Endocrinology Dobesity: relationship between vitamin d deficiency, obesity and sclerostin as a novel biomarker of bone metabolism
JOURNAL OF STEROID BIOCHEMISTRY AND MOLECULAR BIOLOGY Circulating Osteocyte‐Related Biomarkers (vitamin D, sclerostin, dickkopf-1), hepcidin, and oxidative stress markers in early breast cancer: their impact in … Pubmed: 33065276
Oxid Med Cell Longev Long-Term Administration of Abacavir and Etravirine Impairs Semen Quality and Alters Redox System and Bone Metabolism in Growing Male Wistar Rats 34373766
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