ELISA Kit for Alkaline Phosphatase, Tissue-nonspecific (ALPL) Mus musculus (Mouse) Sandwich ELISA

BALP; BSAP; HOPS; AP-TNAP; TNSALP; Bone Alkaline Phosphatase; Alkaline Phosphatase,Tissue-Nonspecific Isozyme; Alkaline Phosphatase, Liver/Bone/Kidney

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  • ELISA Kit for Alkaline Phosphatase, Tissue-nonspecific (ALPL) Packages (Simulation)
  • ELISA Kit for Alkaline Phosphatase, Tissue-nonspecific (ALPL) Packages (Simulation)
  • ELISA Kit for Alkaline Phosphatase, Tissue-nonspecific (ALPL) Results demonstration
  • SEB091Mu.jpg Typical Standard Curve
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 97-105 102
EDTA plasma(n=5) 78-94 83
heparin plasma(n=5) 80-96 87

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Alkaline Phosphatase, Tissue-nonspecific (ALPL) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Alkaline Phosphatase, Tissue-nonspecific (ALPL) 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, Tissue-nonspecific (ALPL) 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) 92-103% 80-101% 96-103% 78-98%
EDTA plasma(n=5) 88-97% 89-96% 80-93% 79-94%
heparin plasma(n=5) 78-94% 80-98% 80-95% 85-102%

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, Tissue-nonspecific (ALPL)

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

Citations

  • Osteopenic bone cell response to strontium-substituted hydroxyapatiteSpringerLink: n8233337w171v451
  • Dextromethorphan inhibits osteoclast differentiation by suppressing RANKL-induced nuclear factor-κB activationPubMed: 23400250
  • Chronic exposure to low concentrations of strontium 90 affects bone physiology but not the hematopoietic system in miceWiley: Source
  • The antidepressant bupropion exerts alleviating properties in an ovariectomized osteoporotic rat modelPubmed:25544359
  • Enhanced differentiation of osteoblastic cells on novel chitosan/β-1, 3-glucan/bioceramic scaffolds for bone tissue regenerationPubmed:25586067
  • Effect of Mirtazapine on Rat Bone Tissue after OrchidectomyPubMed: 25871861
  • New method for the fabrication of highly osteoconductive β-1,3-glucan/HA scaffold for bone tissue engineering: Structural, mechanical, and biological characterization.Pubmed:27239050
  • MiR-142-5p promotes bone repair by maintaining osteoblast activityPubmed:27085967
  • New method for the fabrication of highly osteoconductive β‐1, 3‐glucan/HA scaffold for bone tissue engineering: Structural, mechanical, and biological …doi:10.1002
  • MiR‑142‑5p promotes bone repair by maintaining osteoblast activitypubmed:27085967
  • Hydrolysis of Extracellular Pyrophosphate increases in post-hemodialysis plasmaPubmed:30038263
  • Suppression Effect of Astaxanthin on Osteoclast Formation In Vitro and Bone Loss In VivoPubmed:29562730
  • Effects of Amlodipine on Bone Metabolism in Orchidectomised Spontaneously Hypertensive RatsPubmed:29898457
  • Combination therapy with BMP‑2 and psoralen enhances fracture healing in ovariectomized mice10.3892:etm.2018.6353
  • Long non-coding RNA SNHG7 promotes the fracture repair through negative modulation of miR-9
  • The effect of low temperature atmospheric nitrogen plasma on MC3T3-E1 preosteoblast proliferation and differentiation in vitro
  • Bovine Hydroxyapatite-Based Bone Scaffold with Gentamicin Accelerates Vascularization and Remodeling of Bone Defect34104195

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