MS Mouse LEP(Leptin) ELISA Kit

MS Mouse LEP(Leptin) ELISA Kit

Detection Range	0.63-40 ng/mL
Sensitivity	0.38 ng/mL

Product Details

Properties

Assay type	Sandwich-ELISA
Format	96T
Assay time	3.5h
Detection range	0.63-40 ng/mL
Sensitivity	0.38 ng/mL
Sample type &Sample volume	serum, plasma and other biological fluids; 25μL
Specificity	This kit recognizes Mouse LEP in samples. No significant cross-reactivity or interference between Mouse LEP 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 Mouse LEP 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 Mouse LEP. Standards or samples are added to the micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Mouse LEP 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 Mouse LEP, 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 Mouse LEP. You can calculate the concentration of Mouse LEP in the samples by comparing the OD of the samples to the standard curve.

 

Kit Components and 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 (50×)	96T: 1 vial, 120 μL 48T: 1 vial, 60 μL
Concentrated HRP Conjugate (50×)	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 Mouse LEP were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Mouse LEP 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)	2.13	5.51	15.51	2.27	5.09	16.44
Standard deviation	0.11	0.24	0.79	0.13	0.28	0.61
CV (%)	5.15	4.38	5.12	5.93	5.59	3.73

 

 Recovery

The recovery of Mouse LEP 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)	93-108	100
EDTA plasma (n=8)	93-108	101
Cell culture media (n=8)	90-105	96

Target Information

Assay Procedures

	1. Add 25μL standard or sample to the wells. Incubate for 90 min at 37°C
	2. Discard the liquid, immediately add 50μL 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 50μL HRP conjugate working solution. Incubate for 30 min at 37°C. Aspirate and wash the plate for 5 times
	5. Add 50μL Substrate Reagent. Incubate for 15 min at 37°C
	6. Add 25μL Stop Solution
	7. Read the plate at 450nm immediately. Calculation of the results