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Current Position:Home>>Kits>>ELISA Kits>> Human LEP(Leptin) ELISA Kit

Human LEP(Leptin) ELISA Kit

Citations(27)Uniprot : P41159

  • Cat.No.:E-EL-H6017

  • Reactivity: Human
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  • 96T
  • 48T
  • 24T
  • 96T*5
  • 96T*10
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Product Details

Properties

Assay type Sandwich-ELISA
Format 96T/48T
Assay time 3.5h
Detection range 15.63-1000 pg/mL
Sensitivity 9.38 pg/mL
Sample type &Sample volume serum, plasma and other biological fluids; 100μL
Specificity This kit recognizes Human LEP in samples. No significant cross-reactivity or interference between Human 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 Human 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 Human 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 Human 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 Human 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 Human LEP. You can calculate the concentration of Human LEP in the samples by comparing the OD of the samples to the standard curve.

Kit components & 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.
ItemSpecificationsStorage
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 (100×) 96T: 1 vial, 120 μL
48T: 1 vial, 60 μL
Concentrated HRP Conjugate (100×) 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 Human LEP were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human 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 (pg/mL) 41.50 112.90 459.52 43.08 120.81 467.85
Standard deviation 2.32 5.53 19.21 2.22 5.80 21.47
CV (%) 5.59 4.90 4.18 5.15 4.80 4.59

Recovery

The recovery of Human 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) 91-103 96
EDTA plasma (n=8) 93-104 96
Cell culture media (n=8) 92-100 96

Linearity

Samples were spiked with high concentrations of Human LEP and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.

Target Information

Database LinksSwissProt:   P41159
SynonymsLEPD, OBS, Obesity Homolog
Research AreaCancer, Cardiovascular, Metabolism, Neuroscience, Signal transduction, Stem cells

Assay Procedures

elisa assay procedure 1

1. Add 100μL standard or sample to the wells. Incubate for 90 min at 37°C
elisa assay procedure 2

2. Discard the liquid, immediately add 100μL Biotinylated Detection Ab working solution to each well. Incubate for 60 min at 37°C
elisa assay procedure 3

3. Aspirate and wash the plate for 3 times
elisa assay procedure 4

4. Add 100μL HRP conjugate working solution. Incubate for 30 min at 37°C. Aspirate and wash the plate for 5 times
elisa assay procedure 5

5. Add 90μL Substrate Reagent. Incubate for 15 min at 37°C
elisa assay procedure 6

6. Add 50μL Stop Solution
elisa assay procedure 7

7. Read the plate at 450nm immediately. Calculation of the results

Citations

  1. BIOSENSORS & BIOELECTRONICS (2019) IF: 9.5180
    Porous graphene-black phosphorus nanocomposite modified electrode for detection of leptin

    DOI: 10.1016/j.bios.2019.04.045

    PMID: 31085402
  2. Oncotarget (2016) IF: 5.008
    Adipocytes secreted leptin is a pro-tumor factor for survival of multiple myeloma under chemotherapy

    DOI: 10.18632/oncotarget.13342
    Sample: plasma,adipocytes
  3. ENVIRONMENTAL RESEARCH (2019) IF: 5.026
    Contrasting short-term temperature effects on the profiling of metabolic and stress hormones in non-obese healthy adults: A randomized cross-over trial

    DOI: 10.1016/j.envres.2019.109065

    PMID: 32069742

    		 Sample: Urine
  4. JOURNAL OF ORAL PATHOLOGY & MEDICINE (2019) IF: 4.253
    Elevated leptin levels in temporomandibular joint osteoarthritis promote proinflammatory cytokine IL‐6 expression in synovial fibroblasts

    DOI: 10.1111/jop.12819
    Sample: Cell culture supernatant
  5. Scientific Reports (2021) IF: 4.38
    Obesity-related proteins score as a potential marker of breast cancer risk

    DOI: 10.1038/s41598-021-87583-3

    PMID: 33859244

    		 Sample: Plasma
  6. Scientific Reports (2021) IF: 4.38
    Obesity-related proteins score as a potential marker of breast cancer risk

    DOI: 10.1038/s41598-021-87583-3

    PMID: 33859244

    		 Sample: Plasma
  7. International Journal of Environmental Research and Public Health (2022) IF: 4.614
    Diurnal Nonlinear Recurrence Metrics of Skin Temperature and Their Association with Metabolic Hormones in Contrasting Climate Settings: A Randomized Cross-Over Trial

    DOI: 10.3390/ijerph192215195
    Sample: urine
  8. Frontiers in Immunology (2019) IF: 4.716
    Adiponectin Receptors and Pro-inflammatory Cytokines Are Modulated in Common Variable Immunodeficiency Patients: Correlation With Ig Replacement Therapy

    DOI: 10.3389/fimmu.2019.02812

    PMID: 31827477

    		 Sample: Serum
  9. JOURNAL OF LEUKOCYTE BIOLOGY (2021) IF: 4.962
    Leptin correlates with monocytes activation and severe condition in COVID-19 patients

    DOI: 10.1002/JLB.5HI1020-704R

    PMID: 33404078

    		 Sample: Serum
  10. DIABETES RESEARCH AND CLINICAL PRACTICE (2019) IF: 3.239
    Effect of Ramadan diurnal fasting on visceral adiposity and serum adipokines in overweight and obese individuals

    DOI: 10.1016/j.diabres.2019.05.023

    PMID: 31150725

    		 Sample: Serum
  11. Journal of Clinical Medicine (2020) IF: 3.303
    Association of rs699947 (−2578 C/A) and rs2010963 (−634 G/C) Single Nucleotide Polymorphisms of the VEGF Gene, VEGF-A and Leptin Serum Level, and Cardiovascular Risk in Patients with Excess Body Mass: A Case–Control Study

    DOI: 10.3390/jcm9020469
    Sample: Serum
  12. DRUG AND CHEMICAL TOXICOLOGY (2022) IF: 3.356
    The protective effect of N-acetyl cysteine against selenium toxicity and gamma irradiation in rats

    DOI: 10.1080/01480545.2022.2058010

    PMID: 35361025

    		 Sample: blood
  13. JOURNAL OF DERMATOLOGICAL TREATMENT (2021) IF: 3.359
    Effects of TNF inhibitors and an IL12/23 inhibitor on changes in body weight and adipokine levels in psoriasis patients: A 48-week comparative study

    DOI: 10.1080/09546634.2021.1901845

    PMID: 33704001

    		 Sample: Serum
  14. MEDIATORS OF INFLAMMATION (2019) IF: 3.545
    Adiponectin Expression Is Modulated by Long-Term Physical Activity in Adult Patients Affected by Cystic Fibrosis

    DOI: 10.1155/2019/2153934

    PMID: 31582896

    		 Sample: Serum
  15. JOURNAL OF NEPHROLOGY (2021) IF: 3.902
    Body composition, adipokines, FGF23-Klotho and bone in kidney transplantation: Is there a link?

    DOI: 10.1007/s40620-021-00972-9

    PMID: 33560479

    		 Sample: Serum
  16. SCANDINAVIAN JOURNAL OF GASTROENTEROLOGY (2020) IF: 2.13
    Circadian rhythm abnormalities in patients with inflammatory bowel disease – association with adipokine profile

    DOI: 10.1080/00365521.2020.1737727
    Sample: Serum
  17. Biomed Research International (2018) IF: 2.583
    High Ghrelin Level Predicts the Curve Progression of Adolescent Idiopathic Scoliosis Girls

    DOI: 10.1155/2018/9784083
    Sample: Serum
  18. Lipids in Health and Disease (2019) IF: 2.651
    Leptin, resistin and visfatin as useful predictors of gestational diabetes mellitus

    DOI: 10.1186/s12944-019-1169-2

    PMID: 31836012

    		 Sample: Serum
  19. PLoS One (2017) IF: 2.806
    Placental peptides metabolism and maternal factors as predictors of risk of gestational diabetes in pregnant women. A case-control study

    DOI: 10.1371/journal.pone.0181613
    Sample: Serum
  20. Medicina-Lithuania (2022) IF: 2.948
    Ramadan Intermittent Fasting Is Associated with Changes in Circulating Proprotein Convertase Subtilisin/Kexin Type 9 (PCSK9) in Metabolically Healthy Obese Subjects

    DOI: 10.3390/medicina58040503
    Sample: serum
  21. Personalized Medicine (2017) IF: 1.01
    LEPR gene polymorphism and plasma soluble leptin receptor levels are associated with polycystic ovary syndrome in Han Chinese women

    DOI: 10.2217/pme-2017-0016
    Sample: Plasma
  22. ANZ JOURNAL OF SURGERY (2020) IF: 1.355
    Effect of laparoscopic sleeve gastrectomy on renal function in obese patients

    DOI: 10.1111/ans.15767

    PMID: 32115876

    		 Sample: Serum,Urine
  23. Experimental and Therapeutic Medicine (2019) IF: 1.448
    Correlation of insulin‑resistance with blood fat and glucose in elder patients after surgery for hepatic carcinoma

    DOI: 10.3892/etm.2018.6973
    Sample: Serum
  24. Endokrynologia Polska (2021) IF: 1.582
    New insights into the metabolic-bone crosstalk in active acromegaly

    DOI: 10.5603/EP.a2021.0028
    Sample: serum
  25. Journal of Orthopaedic Surgery and Research (2018) IF: 1.61
    Roles and mechanisms of leptin in osteogenic stimulation in cervical ossification of the posterior longitudinal ligament

    DOI: 10.1186/s13018-018-0864-4
    Sample: Serum
  26. Clinical Hypertension (2020)
    Serum adipocytokines and adiposity as predictive indices of preeclampsia

    DOI: 10.1186/s40885-020-00152-0

    PMID: 33014422

    		 Sample: Serum,Urine
  27. Indian Journal of Medical and Paediatric Oncology (2021)
    Obesity and Breast Cancer: Association of Serum Adiponectin, Leptin, and Adiponectin–Leptin Ratio as Risk Biomarkers

    DOI: 10.4103/ijmpo.ijmpo_203_16
    Sample: Serum
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