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Human PLAP/ALPP(Placental Alkaline Phosphatase) ELISA Kit

Citations(7)Uniprot : P05187

Cat.No.:E-EL-H1976
Reactivity: Human

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Product Details

Properties

Assay type	Sandwich-ELISA
Format	96T/48T
Assay time	3.5h
Detection range	0.16-10 ng/mL
Sensitivity	0.10 ng/mL
Sample type &Sample volume	serum, plasma and other biological fluids; 100μL
Specificity	This kit recognizes Human PLAP/ALPP in samples. No significant cross-reactivity or interference between Human PLAP/ALPP 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 PLAP/ALPP 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 PLAP/ALPP. 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 PLAP/ALPP 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 PLAP/ALPP, 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 PLAP/ALPP. You can calculate the concentration of Human PLAP/ALPP 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 PLAP/ALPP were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human PLAP/ALPP 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 (ng/mL)	0.52	1.03	3.53	0.56	1.04	3.68
Standard deviation	0.03	0.06	0.12	0.04	0.04	0.13
CV (%)	5.77	5.83	3.40	7.14	3.85	3.53

Recovery

The recovery of Human PLAP/ALPP 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)	92-103	97
EDTA plasma (n=8)	86-98	93
Cell culture media (n=8)	93-108	99

Linearity

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

Target Information

Database Links	SwissProt: P05187
Research Area	Cancer, 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

International Journal of Nanomedicine (2019) IF: 4.471
Exosomal microRNA profiling in early and late onset preeclamptic pregnant women reflects pathophysiology

DOI: 10.2147/IJN.S208865

PMID: 31413567
Sample: Plasma
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES (2020) IF: 4.556
Exosomal MicroRNAs in Pregnancy Provides Insight into a Possible Cure for Cancer

DOI: 10.3390/ijms21155384

Sample: Placenta-derived exosomes
CYTOKINE (2019) IF: 3.078
Exosomal Th1/Th2 cytokines in preeclampsia and HIV-positive preeclamptic women on highly active anti-retroviral therapy

DOI: 10.1016/j.cyto.2019.154795

PMID: 31398625
Sample: Serum
PLACENTA (2016) IF: 2.972
Placental exosomes and pre-eclampsia: Maternal circulating levels in normal pregnancies and, early and late onset pre-eclamptic pregnancies

DOI: 10.1016/j.placenta.2016.08.078

Sample: Plasma
European Journal of Obstetrics & Gynecology and Reproductive Biology (2017) IF: 1.666
Placental hypoxia inducible factor -1α & CHOP immuno-histochemical expression relative to maternal circulatory syncytiotrophoblast micro-vesicles in preeclamptic and normotensive pregnancies

DOI: 10.1016/j.ejogrb.2017.11.004

Sample: M,i,c,r,o,v,e,s,i,c,l,e
Journal of Maternal-Fetal & Neonatal Medicine (2017) IF: 1.826
Serum endothelin-1 and placental alkaline phosphatase levels in placenta percreta and normal pregnancies

DOI: 10.1080/14767058.2017.1297412

Sample: Serum
European Journal of Obstetrics & Gynecology and Reproductive Biology (2020) IF: 1.868
The impact of circulating exosomes derived from early and late onset pre-eclamptic pregnancies on inflammatory cytokine secretion by BeWo cells

DOI: 10.1016/j.ejogrb.2020.02.032

PMID: 32114320
Sample: Plasma

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