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Pg(Progesterone) ELISA Kit

Catalog number:E-EL-0154

Size:

96T

Qty:

- +

Price: $495

Reactivity: Universal

Detection Range: 0.31~20 ng/mL

Sensitivity: 0.15 ng/mL

Lead Time: 7~10 daysWelcome to order from local distributors.

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Intended use

This ELISA kit can be applied to the in vitro quantitative determination of Pg concentrations in serum, plasma, urine and saliva.

Test principle

This ELISA kit uses the Competitive-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with Pg. During the reaction, Pg in the sample or standard competes with a fixed amount of Pg on the solid phase supporter for sites on the Biotinylated Detection Ab specific to Pg. 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 the addition of stop solution and the color change is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The concentration of Pg in the samples is then determined by comparing the OD of the samples to the standard curve.

Assay type	Competitive
Format	96T
Assay time	2.0h
Reactivity	Universal
Detection Method	Colormetric
Detection Range	0.31—20 ng/mL
Sensitivity	0.15 ng/mL
Sample Volume	50μL
Sample Type	serum, plasma, urine and saliva

Specificity

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

Typical data

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(ng/mL)	O.D	Average
20	0.099 0.083	0.091
10	0.153 0.131	0.142
5	0.364 0.266	0.315
2.5	0.415 0.869	0.642
1.25	0.908 0.926	0.917
0.62	1.451 1.469	1.46
0.31	2.097 2.191	2.144
0	2.995 2.801	2.898

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Pg were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Pg 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)	0.56	2.22	8.78	0.54	2.01	8.55
Standard deviation	0.05	0.14	0.53	0.05	0.14	0.53
C V (%)	8.39	6.42	5.98	9.35	7.04	6.21

Recovery

The recovery of Pg 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)	80-94	84
EDTA plasma (n=5)	82-96	87
Urine (n=5)	85-109	95

Linearity

Samples were spiked with high concentrations of Pg 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)	Urine (n=5)
1:2	Range (%)	83-90	83-102	84-108
1:2	Average (%)	85	90	93
1:4	Range (%)	80-95	82-95	80-112
1:4	Average (%)	89	89	92
1:8	Range (%)	89-100	81-89	86-96
1:8	Average (%)	92	86	87
1:16	Range (%)	87-102	87-115	84-94
1:16	Average (%)	91	98	88

Kit components & Storage

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

Note: All reagent bottle caps must be tightened to prevent evaporation and microbial pollution.
The volume of reagents in partial shipments is a little more than the volume marked on the label, please use accurate measuring equipment instead of directly pouring into the vial(s).

Other supplies required

Microplate reader with 450 nm wavelength filter
High-precision transfer pipette, EP tubes and disposable pipette tips
Incubator capable of maintaining 37℃
Deionized or distilled water
Absorbent paper
Loading slot for Wash Buffer

Assay Procedure

	1.Add 50 μL standard or sample to each well.
	2.Immediately add 50 μL Biotinylated Detection Ab to each well.
	3.Incubate for 45 min at 37℃. Aspirate and wash 3 times.
	4.Add 100 μL HRP Conjugate to each well. Incubate for 30 min at 37℃. Aspirate and wash 5 times.
	5.Add 90 μL Substrate Reagent. Incubate for 15 min at 37℃.
	6.Add 50 μL Stop Solution.
	7.Read at 450 nm immediately. Calculation of results.

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Verified Customer

Z*ySubmitted [ Jun 01 2019 ]

Species:Human
Sample:serum
Sample source:serum
Detection result:good
Description:This ELISA Kit was used to assay normal human(women and men) serum by us. We operated procedure as Elabscience,s the instruction manual provided and the experimental results were very good------when the human serums were detected without dilution, the testing results were in the range of standard curve, what’s more, It is very near compared with the clinical assay kit! The Pg level of normal men serum is 0.1-3.5 ng/mL and the follicular phase women serum is 0.1-2.1 ng/mL.

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