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No .

E-EL-H0064c

Human ET-1(Endothelin 1) ELISA Kit

Item No:

E-EL-H0064c

Chinese Name:

Alias:

EDN1, ET1, HDLCQ7, PPET1, Preproendothelin-1

Keywords:

ELISA kits

Classification:

One-step (QuicKey Pro) Kit

Retail Price

¥ 1650

Market Price

¥ 1650

This kit is designed for the in vitro quantitative analysis of ET-1 in Human serum, plasma or other biological fluids.

Spec
- 48T
- 96T
- 96T*5

Quantity

Inventory surplus

2997

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Product Comparison

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Describe

Use

This kit is designed for the in vitro quantitative analysis of ET-1 in Human serum, plasma or other biological fluids.

Detection Principle

This kit adopts double antibody sandwich ELISA method. Anti-human ET-1 antibody is used to encapsulate on the enzyme labeling plate. Human ET-1 in the sample or standard will bind to the encapsulated antibody during the experiment, and the free components will be washed away. Biotinylated Anti-Human ET-1 Antibody and horseradish peroxidase labeled affinity are added sequentially. The anti-human ET-1 antibody binds to human ET-1 bound to the coated antibody, the biotin binds specifically to the affinity element and an immune complex is formed, and the free components are washed away. Colorimetric substrate (TMB) was added, TMB showed blue color catalyzed by horseradish peroxidase, and turned into yellow color after addition of termination solution. The OD value was measured at 450 nm with an enzyme marker, and there was a positive ratio between the concentration of ET-1 and the OD450 value, and the concentration of ET-1 in the sample was calculated by plotting the standard curve.

Reaction type	sandwiching method
Specification	96T
Reaction time	3.5h
Reactivity	Human
Detection method	Colormetric
Detection range	1.25—80 pg/mL
Sensitivity	0.75 pg/mL
Sample Volume	100μL
Sample Type	Serum, plasma or other biological fluids

Idiosyncrasy

Detects human ET-1 in samples with no significant cross-reactivity with other related proteins.

Repeatable

The coefficients of variation within plates and between plates were all <10%.

Typical data

The OD values of the standard curve will vary due to different experimental operating conditions (e.g., operator, pipetting technique, plate washing technique and stabilization conditions, etc.). The following data and curves are for reference only, and the experimenter needs to establish the standard curve according to his/her own experiment.

(pg/mL)	O.D	Average	Corrected
80	2.445 2.501	2.473	2.394
40	1.632 1.638	1.635	1.556
20	0.902 0.896	0.899	0.82
10	0.466 0.492	0.479	0.4
5	0.273 0.243	0.258	0.179
2.5	0.185 0.173	0.179	0.1
1.25	0.121 0.139	0.13	0.051
0	0.076 0.082	0.079	--

Precision

Intra-plate precision: Low, medium and high concentration samples were tested 20 times on 1 plate.
　　Inter-plate precision: low concentration, medium concentration and high concentration samples were tested 20 times on 3 plates.

	Intra-assay Precision			Inter-assay Precision
Sample	1	2	3	1	2	3
n	20	20	20	20	20	20
Mean (pg/mL)	4.00	7.80	33.60	4.00	7.70	36.30
Standard deviation	0.30	0.40	1.40	0.20	0.30	1.40
C V (%)	7.50	5.13	4.17	5.00	3.90	3.86

Recovery rate

A known concentration of the target protein was added to five different samples and recovery experiments were done to obtain a range of recoveries and an average recovery.

Sample Type	Range (%)	Average Recovery (%)
Serum (n=8)	92-103	97
EDTA plasma (n=8)	89-104	96
Cell culture media (n=8)	86-100	92

Linearity

A recovery experiment was performed by adding known concentrations of target protein to 5 samples to obtain a range of recoveries and average recoveries. Dilute the 5 samples 2x, 4x, 8x and 16x to obtain the recovery range and average recovery.

		Serum (n=5)	EDTA plasma (n=5)	Cell culture media (n=5)
1:2	Range (%)	89-104	86-97	93-110
1:2	Average (%)	95	91	100
1:4	Range (%)	95-110	87-97	92-104
1:4	Average (%)	101	92	99
1:8	Range (%)	95-105	84-95	96-111
1:8	Average (%)	100	89	103
1:16	Range (%)	99-115	82-93	96-109
1:16	Average (%)	105	88	102

Kit Composition and Storage

Unopened kits can be stored at 2-8°C for one week; if the kit is to be used after one week, unpack the kit and store the components separately according to the conditions in the table below.

Chinese name	English name	Specification	Storage conditions
ELISA酶标板(可拆卸)	Micro ELISA Plate(Dismountable)	8 holes×12 strips	-20℃, can be stored for 6 months
冻干标准品	Reference Standard	2 sticks
浓缩生物素化抗体(100×)	Concentrated Biotinylated Detection Ab	1 x 120 μL
浓缩HRP酶结合物(100×)	Concentrated HRP Conjugate	1 x 120 μL	-20℃ (avoid light), can be stored for 6 months
标准品&样品稀释液	Reference Standard & Sample Diluent	1 bottle 20 mL	2-8℃. Can be stored for 6 months
生物素化抗体稀释液	Biotinylated Detection Ab Diluent	1 bottle 14 mL
酶结合物稀释液	HRP Conjugate Diluent	1 bottle 14 mL
浓缩洗涤液(25×)	Concentrated Wash Buffer (25×)	1 bottle 30 mL
底物溶液(TMB)	Substrate Reagent	1 bottle 10 mL	2-8℃(avoid light)
反应终止液	Stop Solution	1 bottle 10 mL	2-8℃
封板覆膜	Plate Sealer	5 sheets
产品说明书	Product Description	1 sheet
质检报告	Certificate of Analysis	1 sheet

Note: All reagent bottle caps must be screwed on tightly to prevent evaporation and microbial contamination.
Reagent volumes are based on the actual shipping version of the instructions. Reagents may be dispensed in a slightly larger volume than indicated on the label, so please measure rather than pour when using.

Self-contained items required for testing

•1. Enzyme Labeling Instrument (450nm wavelength filter)

•2. High-precision pipettes, EP tubes and disposable tips: 0.5-10 μL, 2-20 μL, 20-200 μL, 200-1000 μL

•3.37℃ thermostat, double-distilled water or deionized water

•4. Pipette

Procedure

	1. Add 100μL of standard working solution or sample into the corresponding plate wells and incubate at 37℃ for 90 minutes.
	2. Discard the liquid in the plate, immediately add 100μL of biotinylated antibody working solution and incubate at 37℃ for 60 minutes.
	3. Discard the liquid in the plate and wash the plate 3 times.
	4. add 100μL of HRP enzyme conjugate working solution to each well, incubate at 37℃ for 30 minutes, discard the liquid in the plate, and wash the plate five times.
	5. Add 90μL of substrate solution to each well and incubate at 37℃ for 15 minutes.
	6. Add 50μL of termination solution to each well.
	7. Immediately read at 450nm and process the data.