LEP Mini-sample ELISA kit (Human)

SKU: MEA084Hu

Size: 96Tests

# of Times Cited in literature: -

Prepare Time: 2-7 days(please inquire for mutiple units)

Target Name: LEP

Target Full Name: Leptin

Alternative Names: OB; OBS; Obesity Homolog; Obesity Factor; Obese Protein

Target Species: Human

Uniprot: P41159

Gene ID: 3952

Featured Series: ME kit

Featured Series Function: Detects protein (micro-volume samples 15-25 μl)

Specificity: Reactive with Human LEP / Leptin

Method: Colormetric

Detection principle: Double-antibody Sandwich

Detection range: 0.156-10ng/mL

Sensitivity: 0.056ng/mL

Assay Time: 3h

Sample Size: 15-25uL

Recommended/Predicted Sample Types: Serum, Plasma, Saliva, Tissue Homogenates, Cell Lysates, Cell Culture Supernates and other Biological Fluids

Assay Precision: Intra-Assay: CV<10%, Inter-Assay: CV<12%

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

Storage: 4°C for 1 month/ -20°C for long-term(One year within shelf life)

Shelf-life: 12 months

Specificity: This assay has high sensitivity and excellent specificity for detection of Mini Samples Leptin (LEP). No significant cross-reactivity or interference between Mini Samples Leptin (LEP) and analogues was observed.

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.

Assay procedure summary: 1. Prepare all reagents, samples and standards; 2. Add 25µL standard or sample to each well. Incubate 1 hour at 37°C; 3. Aspirate and add 25µL prepared Detection Reagent A. Incubate 1 hour at 37°C; 4. Aspirate and wash 3 times; 5. Add 25µL prepared Detection Reagent B. Incubate 30 minutes at 37°C; 6. Aspirate and wash 5 times; 7. Add 25µL Substrate Solution. Incubate 10-20 minutes at 37°C; 8. Add 20µL Stop Solution. Read at 450nm immediately.

Test principle: The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Mini Samples Leptin (LEP). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Mini Samples Leptin (LEP). Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Mini Samples Leptin (LEP), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of Mini Samples Leptin (LEP) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Research Area: Metabolic pathway;Endocrinology;Cardiovascular biology;

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