ELISA Kit for Sclerostin (SOST)

VBCH; Sclerosteosis

Specificity

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

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 97-104 101
EDTA plasma(n=5) 85-101 90
heparin plasma(n=5) 83-101 88

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Sclerostin (SOST) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Sclerostin (SOST) 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) 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) 78-105% 80-103% 79-103% 87-94%
EDTA plasma(n=5) 92-103% 79-93% 96-105% 98-105%
heparin plasma(n=5) 81-92% 87-94% 87-103% 89-97%

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
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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