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ELISA Kit for Alkaline Phosphatase (ALP) Bos taurus; Bovine (Cattle) Sandwich ELISA

AKP; ALKP; Basic Phosphatase

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  • ELISA Kit for Alkaline Phosphatase (ALP) Packages (Simulation)
  • ELISA Kit for Alkaline Phosphatase (ALP) Packages (Simulation)
  • ELISA Kit for Alkaline Phosphatase (ALP) Results demonstration
  • SEB472Bo.jpg Typical Standard Curve
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 96-105 101
EDTA plasma(n=5) 87-95 91
heparin plasma(n=5) 93-101 97

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Alkaline Phosphatase (ALP) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Alkaline Phosphatase (ALP) 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 Alkaline Phosphatase (ALP) 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) 85-103% 80-101% 81-93% 83-92%
EDTA plasma(n=5) 78-95% 89-96% 79-89% 87-101%
heparin plasma(n=5) 91-101% 92-102% 78-97% 78-95%

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 Alkaline Phosphatase (ALP)

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

Citations

  • Comparison of platelet rich fibrin and collagen as osteoblast-seeded scaffolds for bone tissue engineering applicationsWiley: source
  • The role of atorvastatin in bone metabolism in male albino Wistar ratsIngenta: art00010
  • Combined effect of strontium and zoledronate on hydroxyapatite structure and bone cell responsesScienceDirect: S0142961214003135
  • Directing chondrogenic differentiation of mesenchymal stem cells with a solid-supported chitosan thermogel for cartilage tissue engineeringPubmed:24770944
  • Strontium and zoledronate hydroxyapatites graded composite coatings for bone prosthesesPubmed:25706198
  • The active role of osteoporosis in the interaction between osteoblasts and bone metastasesPubMed: 26057367
  • Multi‐Layered Scaffolds for Osteochondral Tissue Engineering: In Vitro Response of Co‐Cultured Human Mesenchymal Stem CellsPubMed: 26126665
  • Biological Evaluation of a Prototype Material made of Polyglycolic Acid and HydroxyapatiteArticle: Jhtb
  • Antiresorption implant coatings based on calcium alendronate and octacalcium phosphate deposited by matrix assisted pulsed laser evaporationPubMed: 26445021
  • On the Mechanism of Drug Release from Polysaccharide Hydrogels Cross-Linked with Magnetite Nanoparticles by Applying Alternating Magnetic Fields: the Case of DOXO Delivery2310-2861: 1
  • Antioxidant and bone repair properties of quercetin-functionalized hydroxyapatite: an in vitro osteoblast-osteoclast-endothelial cell co-culture studyPubMed: 26689470
  • A composite scaffold of MSC affinity peptide-modified demineralized bone matrix particles and chitosan hydrogel for cartilage regenerationPubMed: 26632447
  • Antioxidant and bone repair properties of quercetin-functionalized hydroxyapatite: An in vitro osteoblast–osteoclast–endothelial cell co-culture studyPubmed:26689470
  • Extracorporeal shock waves alone or combined with raloxifene promote bone formation and suppress resorption in ovariectomized rats.pubmed:28158228
  • Adiponectin protects the rats liver against chronic intermittent hypoxia induced injury throughAMP-activated protein kinase pathway.pubmed:27678302
  • An advanced tri-culture model to evaluate the dynamic interplay among osteoblasts, osteoclasts, and endothelial cells.pubmed:28240358

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