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Human ALPL(Alkaline Phosphatase, Liver/Bone/Kidney) ELISA Kit

Citations(3)Uniprot : P05186

Cat.No.:E-EL-H2598
Reactivity: Human

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Product Details

Properties

Assay type	Sandwich-ELISA
Format	96T/48T
Assay time	3.5h
Detection range	78.13-5000 pg/mL
Sensitivity	46.88 pg/mL
Sample type &Sample volume	serum, plasma and other biological fluids; 100μL
Specificity	This kit recognizes Human ALPL in samples. No significant cross-reactivity or interference between Human ALPL and analogues was observed.
Reproducibility	Both intra-CV and inter-CV are < 10%.
Application	This ELISA kit applies to the in vitro quantitative determination of Human ALPL concentrations in serum, plasma and other biological fluids.

Test Principle

This ELISA kit uses the Sandwich-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Human ALPL. Standards or samples are added to the micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Human ALPL and Avidin-Horseradish Peroxidase (HRP) conjugate are added successively to each micro plate well and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Human ALPL, biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by the addition of stop solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The OD value is proportional to the concentration of Human ALPL. You can calculate the concentration of Human ALPL in the samples by comparing the OD of the samples to the standard curve.

Kit components & Storage

An unopened kit can be stored at 2-8℃ for 1 month. If the kit is not supposed to be used within 1 month, store the components separately according to the following conditions once the kit is received.

Item	Specifications	Storage
Micro ELISA Plate(Dismountable)	96T: 8 wells ×12 strips 48T: 8 wells ×6 strips	-20℃, 6 months
Reference Standard	96T: 2 vials 48T: 1 vial
Concentrated Biotinylated Detection Ab (100×)	96T: 1 vial, 120 μL 48T: 1 vial, 60 μL
Concentrated HRP Conjugate (100×)	96T: 1 vial, 120 μL 48T: 1 vial, 60 μL	-20℃(Protect from light), 6 months
Reference Standard & Sample Diluent	1 vial, 20 mL	2-8°C, 6 months
Biotinylated Detection Ab Diluent	1 vial, 14 mL
HRP Conjugate Diluent	1 vial, 14 mL
Concentrated Wash Buffer (25×)	1 vial, 30 mL
Substrate Reagent	1 vial, 10 mL	2-8℃(Protect from light)
Stop Solution	1 vial, 10 mL	2-8°C
Plate Sealer	5 pieces
Manual	1 copy
Certificate of Analysis	1 copy

Technical Data

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human ALPL were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human ALPL were tested on 3 different plates, 20 replicates in each plate.

	Intra-assay Precision			Inter-assay Precision
Sample	1	2	3	1	2	3
n	20	20	20	20	20	20
Mean (pg/mL)	232.41	686.93	2152.29	235.06	692.44	2121.00
Standard deviation	15.27	28.64	98.79	13.26	38.85	75.51
CV (%)	6.57	4.17	4.59	5.64	5.61	3.56

Recovery

The recovery of Human ALPL spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.

Sample Type	Range (%)	Average Recovery (%)
Serum(n=8)	96-110	102
EDTA plasma (n=8)	87-103	94
Cell culture media (n=8)	94-105	100

Linearity

Samples were spiked with high concentrations of Human ALPL and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.

Target Information

Database Links	SwissProt: P05186
Research Area	Cancer, Cell Biology, Tags and Cell Markers

Assay Procedures

	1. Add 100μL standard or sample to the wells. Incubate for 90 min at 37°C
	2. Discard the liquid, immediately add 100μL Biotinylated Detection Ab working solution to each well. Incubate for 60 min at 37°C
	3. Aspirate and wash the plate for 3 times
	4. Add 100μL HRP conjugate working solution. Incubate for 30 min at 37°C. Aspirate and wash the plate for 5 times
	5. Add 90μL Substrate Reagent. Incubate for 15 min at 37°C
	6. Add 50μL Stop Solution
	7. Read the plate at 450nm immediately. Calculation of the results

Citations

BONE (2019) IF: 4.36
Daily oligofructose-enriched inulin intake impacts bone turnover markers but not the cytokine profile in pediatric patients with celiac disease on a gluten-free diet: Results of a randomised, placebo-controlled pilot study

DOI: 10.1016/j.bone.2019.03.001

Sample: Plasma
AMERICAN JOURNAL OF REPRODUCTIVE IMMUNOLOGY (2017) IF: 3.013
Analysis of the ectoenzymes ADA, ALP, ENPP1, and ENPP3, in the contents of ovarian endometriomas as candidate biomarkers of endometriosis

DOI: 10.1111/aji.12794

Sample: Tissue homogenate
Applied Sciences-Basel (2022) IF: 2.838
Alendronate Prevents Early Periprosthetic Bone Loss in Cementless Total Hip Arthroplasty Better Than Simvastatin

DOI: 10.3390/app12168054

Sample: serum

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Reviews/Q&A

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Q&A

Z****************** Submitted [ Jan 12 2023 ]

Asked: Hi. I am a postgraduate dental student from Kenya and I wish to conduct a study for my thesis using SALIVA for detection of Alkaline Phosphatase enzyme. Can the ELISA kit be used for this purpose? Please advise. Thanks.

adminSubmitted [ Jan 12 2023 ]

Answered: Dear Zun-noorain Noorani, Thanks for reaching out Elabscience. Regarding your inquired product Human ALPL(Alkaline Phosphatase, Liver/Bone/Kidney) ELISA Kit, #E-EL-H2598, we tested the kit using human saliva samples before, the reference values were below 46pg/mL, therefore the kit may be not suitable for human saliva samples. Please let us know if we could be of any further assistance.

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