8-epi-PGF2alpha(8-Epi-Prostaglandin F2 Alpha) ELISA Kit

Mfr.No.

E-EL-0041

Antigen

8-epi-PGF2α

Alternative Names

15-F2t-IsoP ; 8-iso-PGF2a ; 8-isoprostane ; 8-iso-PG ; 8-iso Prostaglandin

Background

This kit uses competitive-ELISA as the method. The micro ELISA plate provided in this kit has been pre-coated with ; 8-epi-PGF2α .During the reaction, 8-epi-PGF2α in the sample or standard competes with a fixed amount of 8-epi-PGF2α on the solid phase supporter for sites on the Biotinylated Detection Ab specific to 8-epi-PGF2α Excess conjugate and unbound sample or standard are washed from the plate, and Avidin conjugated to Horseradish Peroxidase (HRP) are added to each microplate well and incubated. Then a TMB substrate solution is added to each well. The enzyme-substrate reaction is terminated by adding Stop Solution and the color change is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The concentration of 8-epi-PGF2α in the samples is then determined by comparing the OD of the samples to the standard curve.

Description

As the OD values of the standard curve may vary according to the conditions of the actual assay performance (e.g. operator, pipetting technique, washing technique or temperature effects), the operator should establish a standard curve for each test. Typical standard curve and data is provided below for reference only.

Concentration(pg/mL)	O.D	Average
1000	0.334 0.356	0.345
500	0.422 0.46	0.441
250	0.636 0.604	0.62
125	0.907 0.9330000000000001	0.92
62.5	1.347 1.333	1.34
31.25	1.806 1.8	1.803
15.63	2.188 2.204	2.196
0	2.773 2.775	2.774

Specificity

This kit recognizes 8-epi-PGF2α in samples. No significant cross-reactivity or interference between 8-epi-PGF2α and analogues was observed.

Assay Time

2.5h

Components

An unopened kit can be stored at 4℃ for 1 month. If the kit is not used within 1 month, store the items separately according to the following conditions once the kit is received.

Item	Specifications	Storage
Micro ELISA Plate(Dismountable)	8 wells ×12 strips	-20℃, 6 months
Reference Standard	2 vials
Concentrated Biotinylated Detection Ab (100×)	1 vial, 120 μL
Concentrated HRP Conjugate (100×)	1 vial, 120 μL	-20℃(shading light), 6 months
Reference Standard & Sample Diluent	1 vial, 20 mL	4℃, 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	4℃(shading light)
Stop Solution	1 vial, 10 mL	4℃
Plate Sealer	5 pieces
Product Description	1 copy
Certificate of Analysis	1 copy

Detection Method

Colormetric

Detection Range

15.625--1000pg/mL

Linearity

Samples were spiked with high concentrations of 8-epi-PGF2α and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.

		Serum (n=5)	EDTA plasma (n=5)	Cell culture media (n=5)
1:2	Range (%)	86-97	87-102	88-99
1:2	Average (%)	91	94	94
1:4	Range (%)	83-95	87-100	101
1:4	Average (%)	90	93	101
1:8	Range (%)	85-101	88-101	98-114
1:8	Average (%)	92	95	105
1:16	Range (%)	83-94	85-99	99-114
1:16	Average (%)	89	92	105

Method Type

Competitive

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level 8-epi-PGF2α were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level 8-epi-PGF2α 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)	51.20	101.80	446.40	54.50	95.70	463.70
Standard deviation	2.90	4.50	15.60	3.50	5.00	25.00
C V (%)	5.66	4.42	3.49	6.42	5.22	5.39

Recovery

The recovery of 8-epi-PGF2α 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=5)	85-97	92
EDTA plasma (n=5)	92-108	100
Cell culture media (n=5)	93-107	100

Sample Type

Serum, Plasma, Cell supernatant

Sensitivity

9.38 pg/mL

Manual

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SKU	Supplier No.	Size	Your price
BHE10302522	E-EL-0041	96T	$495	Add to Cart
BHE10302522	E-EL-0041	96T*5	$2227.50	Add to Cart
BHE10302522	E-EL-0041	96T*10	$4207.50	Add to Cart
BHE10302522	E-EL-0041	48T	$396	Add to Cart

8-epi-PGF2alpha(8-Epi-Prostaglandin F2 Alpha) ELISA Kit

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