ELISA Kit for Osteocalcin (OC)
FOR IN VITRO AND RESEARCH USE ONLY, NOT FOR USE IN CLINICAL DIAGNOSTIC PROCEDURES!
Organism species Rattus norvegicus (Rat)
Product No.SEA471Ra
Sample typeSerum, plasma, tissue homogenates, cell lysates and other biological fluids.
Format96-well strip plate
Assay length4.5 hours
Detection range15.63-1000pg/mL The standard curve concentrations used for the ELISA’s were 1000pg/mL, 500pg/mL, 250pg/mL, 125pg/mL, 62.5pg/mL, 31.25pg/mL, 15.63pg/mL
SensitivityThe minimum detectable dose of this kit is typically less than 6.3pg/mL.
Specificity
This assay has high sensitivity and excellent specificity for detection of Osteocalcin (OC).
No significant cross-reactivity or interference between Osteocalcin (OC) and analogues was observed.
Recovery
Matrices listed below were spiked with certain level of recombinant Osteocalcin (OC) and the recovery rates were calculated by comparing the measured value to the expected amount of Osteocalcin (OC) in samples.
Matrix Recovery range (%) Average(%)
serum(n=5)86-10294
EDTA plasma(n=5)96-105101
heparin plasma(n=5)83-9894
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Osteocalcin (OC) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Osteocalcin (OC) 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 Osteocalcin (OC) 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)82-93%86-94%79-97%78-91%
EDTA plasma(n=5)81-105%81-101%89-96%93-101%
heparin plasma(n=5)94-102%84-101%93-101%86-97%
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 2 hours at 37oC;
3. Aspirate and add 100µL prepared Detection Reagent A. Incubate 1 hour at 37oC;
4. Aspirate and wash 3 times;
5. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37oC;
6. Aspirate and wash 5 times;
7. Add 90µL Substrate Solution. Incubate 15-25 minutes at 37oC;
8. Add 50µL Stop Solution. Read at 450nm immediately.
Test principle
The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Osteocalcin (OC). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Osteocalcin (OC). Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Osteocalcin (OC), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of Osteocalcin (OC) in the samples is then determined by comparing the O.D. of the samples to the standard curve.
Package and Components
Reagent Preparation
Results demonstration
Typical Standard Curve
Certificate
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Free use and Guarantees
Free trial
In case it is difficult to pre-evaluate experiment result because of specific test samples or other reasons, free trial can be offered.
If the budget is really limited or other special situation, you can apply the free products.
Guarantees
Unconditionally return policy within 72 hours.
For any complaint, we promise to reply within one work day, resolve the problem within three work days. Implementing initial consultation system in order to provide satisfactory service for all the customers. Customers can unconditionally to request a charge back or refund.
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MagazineReference
Annals of RSCB Biochemical study of the influence of ad type biophytomodulators in healing of bone defects in rats.    Read more(Analelesnbc: source)
Archives of Oral Biology Oxytocin promotes bone formation during the alveolar healing process in old acyclic female rats    Read more(ScienceDirect: S0003996912001021)
Life Sciences Exendin-4 increases bone mineral density in type 2 diabetic OLETF rats potentially through the down-regulation of SOST/sclerostin in osteocytes    Read more(ScienceDirect: S0024320513000027)
Romanian Journal of Morphology and Embryology Serum changes induced by intramedullar experimental administration of bisphosphonates    Read more(PubMed: 21424089)
Nutr Hosp. Effect of the “protein diet” and bone tissue.    Read more(Pubmed: 24483972)
Catalog No. Related products for research use of Rattus norvegicus (Rat) Organism species Applications
MAA471Ra21Monoclonal Antibody to Osteocalcin (OC)WB, ICC, IHC-P, IHC-F, ELISA
PAA471Ra01Polyclonal Antibody to Osteocalcin (OC)WB, ICC, IHC-P, IHC-F, ELISA
PAA471Ra71Biotin-Linked Antibody to Osteocalcin (OC)IHC;WB;ELISA
PAA471Ra81FITC-Linked Antibody to Osteocalcin (OC)IHC;ICC;IF;WB
RPA471Ra01Recombinant Osteocalcin (OC)SDS-PAGE; WB; ELISA; IP.
SEA471RaELISA Kit for Osteocalcin (OC)Enzyme-linked immunosorbent assay