CLIA Kit for Leptin (LEP)Homo sapiens (Human)Sandwich CLIA

OB; OBS; Obesity Homolog; Obesity Factor; Obese Protein

  • Product No.SCA084Hu
  • Organism SpeciesHomo sapiens (Human)Same name, Different species.
  • Sample TypeSerum, plasma, saliva, tissue homogenates, cell lysates, cell culture supernates and other biological fluids
  • Test MethodDouble-antibody Sandwich
  • Assay Length2h, 40min
  • Detection Range468.75-30000pg/mL
  • SensitivityThe minimum detectable dose of this kit is typically less than 15.1pg/mL.
  • Specificity This assay has high sensitivity and excellent specificity for detection of Leptin (LEP).
    No significant cross-reactivity or interference between Leptin (LEP) and analogues was observed.
  • ApplicationsChemiluminescent immunoassay for Antigen Detection
  • DownloadInstruction Manual
  • Format48T96T96T*596T*1096T*100
  • FOBUS$ 588US$ 840US$ 3780US$ 7140US$ 58800
  • CLIA Kit for Leptin (LEP)Package and Components
  • CLIA Kit for Leptin (LEP)Package and Components
  • CLIA Kit for Leptin (LEP)Results demonstration
  • SCA084Hu.jpgTypical Standard Curve
  • CertificateISO9001: 2008, ISO13485: 2003 Registered

Recovery

Matrices listed below were spiked with certain level of recombinant Leptin (LEP) and the recovery rates were calculated by comparing the measured value to the expected amount of Leptin (LEP) in samples.

MatrixRecovery range (%)Average(%)
serum(n=5)99-105102
EDTA plasma(n=5)88-9692
heparin plasma(n=5)80-10197

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Leptin (LEP) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Leptin (LEP) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Leptin (LEP) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

Sample1:21:41:81:16
serum(n=5)80-103%84-104%93-103%85-99%
EDTA plasma(n=5)95-104%98-105%97-105%84-98%
heparin plasma(n=5)82-101%79-101%83-91%87-95%

Stability

The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.

Reagents and materials provided

ReagentsQuantityReagentsQuantity
Pre-coated, ready to use 96-well strip plate1Plate sealer for 96 wells4
Standard2Standard Diluent1×20mL
Detection Reagent A1×120µLAssay Diluent A1×12mL
Detection Reagent B1×120µLAssay Diluent B1×12mL
Substrate A1×10mLSubstrate B1×2mL
Wash Buffer (30 × concentrate)1×20mLInstruction manual1

Assay procedure summary

1. Prepare all reagents, samples and standards;
2. Add 100µL standard or sample to each well. Incubate 1 hours at 37°C;
3. Aspirate and add 100µL prepared Detection Reagent A. Incubate 1 hour at 37°C;
4. Aspirate and wash 3 times;
5. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
6. Aspirate and wash 5 times;
7. Add 100µL Substrate Solution. Incubate 10 minutes at 37°C;
8. Read RLU value immediately.

CLIA Kit for Leptin (LEP)

Test principle

The microtiter plate provided in this kit has been pre-coated with an antigen. Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated secondary antibody. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. Then the mixture of substrate A and B is added to generate glow light emission kinetics. Upon plate development, the intensity of the emitted light is proportional to the Leptin (LEP) level in the sample or standard.

Reference

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