ELISA Kit for Acid Phosphatase 5, Tartrate Resistant (ACP5) Rattus norvegicus (Rat) Sandwich ELISA

TRAP; TrATPase; Tartrate Resistant Acid Phosphatase; Type 5 acid phosphatase

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  • ELISA Kit for Acid Phosphatase 5, Tartrate Resistant (ACP5) Packages (Simulation)
  • ELISA Kit for Acid Phosphatase 5, Tartrate Resistant (ACP5) Packages (Simulation)
  • ELISA Kit for Acid Phosphatase 5, Tartrate Resistant (ACP5) Results demonstration
  • SEA902Ra.jpg Typical Standard Curve
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

Recovery

Matrices listed below were spiked with certain level of recombinant Acid Phosphatase 5, Tartrate Resistant (ACP5) and the recovery rates were calculated by comparing the measured value to the expected amount of Acid Phosphatase 5, Tartrate Resistant (ACP5) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 79-103 84
EDTA plasma(n=5) 91-104 95
heparin plasma(n=5) 89-97 93

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Acid Phosphatase 5, Tartrate Resistant (ACP5) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Acid Phosphatase 5, Tartrate Resistant (ACP5) 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 Acid Phosphatase 5, Tartrate Resistant (ACP5) 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) 93-105% 80-104% 94-102% 86-95%
EDTA plasma(n=5) 82-97% 86-101% 84-98% 80-102%
heparin plasma(n=5) 78-93% 80-89% 97-105% 81-94%

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.

ELISA Kit for Acid Phosphatase 5, Tartrate Resistant (ACP5)

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 Acid Phosphatase 5, Tartrate Resistant (ACP5). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Acid Phosphatase 5, Tartrate Resistant (ACP5). 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 Acid Phosphatase 5, Tartrate Resistant (ACP5), 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 Acid Phosphatase 5, Tartrate Resistant (ACP5) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Citations

  • Oxytocin promotes bone formation during the alveolar healing process in old acyclic female ratsScienceDirect: S0003996912001021
  • Exendin-4 Increases Bone Mineral Density in Type 2 Diabetic OLETF Rats Potentially Through the Down-Regulation of SOST/Sclerostin in OsteocytesPubMed: 23357248
  • Valvular osteoclasts in calcification and aortic valve stenosis severity.Pubmed: 23452891
  • Antiosteoporosis Activity of New Oriental Medicine Preparation (Kyungokgo Mixed with Water Extract of Hovenia dulcis) on the Ovariectomized MicePubmed:25737735
  • The influence of bexarotene, a selective agonist of the retinoid receptor X (RXR), and tazarotene, a selective agonist of the retinoid acid receptor (RAR), on bone metabolism in ratsPubMed: 26569440
  • The effects of strength training and raloxifene on bone health in aging ovariectomized ratsPubmed:26812611
  • In Vitro and In Vivo Effects of Gracilaria verrucosa Extracts on Osteoclast Differentiation.pubmed:28335442

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