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6-keto-PGF1a(6-keto-prostaglandin F1a) ELISA Kit

Cat.No.:E-EL-0054
Reactivity: Universal

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96T
48T
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96T*5
96T*10

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Product Details

Properties

Assay type	Competitive-ELISA
Format	96T/48T
Assay time	2.0h
Detection range	15.63-1000 pg/mL
Sensitivity	9.38 pg/mL
Sample type &Sample volume	serum, plasma and other biological fluids; 50μL
Specificity	This kit recognizes 6-keto-PGF1a in samples. No significant cross-reactivity or interference between 6-keto-PGF1a 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 6-keto-PGF1a concentrations in serum, plasma and other biological fluids.

Test Principle

This ELISA kit uses the Competitive-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with 6-keto-PGF1a. During the reaction, 6-keto-PGF1a in the sample or standard competes with a fixed amount of 6-keto-PGF1a on the solid phase supporter for sites on the Biotinylated Detection Ab specific to 6-keto-PGF1a. Excess conjugate and unbound sample or standard are washed away, and Avidin-Horseradish Peroxidase (HRP) conjugate are added to each micro plate well and incubated. Then a TMB substrate solution is added to each well. The enzyme-substrate reaction is terminated by the addition of stop solution and the color turns from blue to yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The concentration of 6-keto-PGF1a in tested samples can be calculated 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℃
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 6-keto-PGF1a were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level 6-keto-PGF1a 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)	54.40	139.60	366.20	50.50	151.20	402.50
Standard deviation	3.60	5.70	17.90	3.00	8.00	14.90
CV (%)	6.62	4.08	4.89	5.94	5.29	3.70

Recovery

The recovery of 6-keto-PGF1a 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)	91-104	96
EDTA plasma(n=8)	83-97	90
Cell culture media(n=8)	87-103	94

Linearity

Samples were spiked with high concentrations of 6-keto-PGF1a and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.

Target Information

Research Area

Cardiovascular

Assay Procedures

	1. Add 50μL standard or sample to the wells, immediately add 50μL Biotinylated Detection Ab working solution to each well. Incubate for 45 min at 37°C
	2. Aspirate and wash the plate for 3 times
	3. Add 100μL HRP conjugate working solution. Incubate for 30 min at 37°C. Aspirate and wash the plate for 5 times
	4. Add 90μL Substrate Reagent. Incubate for 15 min at 37°C
	5. Add 50μL Stop Solution
	6. Read the plate at 450nm immediately. Calculation of the results

Citations

Theranostics (2019) IF: 8.0630
Temporal inhibition of mouse mammary gland cancer metastasis by CORM-A1 and DETA/NO combination therapy

DOI: 10.7150/thno.31461

Sample: plasma
BIOMEDICINE & PHARMACOTHERAPY (2023) IF: 7.419
Quince extract resists atherosclerosis in rats by down-regulating the EGFR/PI3K/Akt/GSK-3β pathway

DOI: 10.1016/j.biopha.2023.114330

Sample: serum
Chinese Medicine (2021) IF: 5.455
Anti-platelet aggregation of Panax notoginseng triol saponins by regulating GP1BA for ischemic stroke therapy

DOI: 10.1186/s13020-021-00424-3

PMID: 33468191
Sample: Serum
Evidence-based Complementary and Alternative Medicine (2021) IF: 2.63
The Anti-Inflammatory Effect of Smilax china L. Extract on LPS-Stimulated THP-1 via Downregulation of MAPK and NF-κB Signaling Pathway

DOI: 10.1155/2021/9958808

Sample: THP-1 cells
ACS Omega (2020) IF: 2.87
Extract from Rostellularia procumbens (L.) Nees Inhibits Thrombosis and Platelet Aggregation by Regulating Integrin β3 and MAPK Pathways

DOI: 10.1021/acsomega.0c05227

Sample: serum

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