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

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

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  • ELISA Kit for Leptin (LEP) Packages (Simulation)
  • ELISA Kit for Leptin (LEP) Packages (Simulation)
  • ELISA Kit for Leptin (LEP) Results demonstration
  • SEA084Hu.jpg Typical Standard Curve
  • Certificate ISO9001: 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.

Matrix Recovery range (%) Average(%)
serum(n=5) 97-104 101
EDTA plasma(n=5) 78-90 82
heparin plasma(n=5) 88-104 94

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.

Sample 1:2 1:4 1:8 1:16
serum(n=5) 94-102% 86-101% 99-105% 86-94%
EDTA plasma(n=5) 80-101% 91-101% 90-99% 91-101%
heparin plasma(n=5) 79-101% 85-102% 79-101% 96-105%

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

Reagents Quantity Reagents Quantity
Pre-coated, ready to use 96-well strip plate 1 Plate sealer for 96 wells 4
Standard 2 Standard Diluent 1×20mL
Detection Reagent A 1×120µL Assay Diluent A 1×12mL
Detection Reagent B 1×120µL Assay Diluent B 1×12mL
TMB Substrate 1×9mL Stop Solution 1×6mL
Wash Buffer (30 × concentrate) 1×20mL Instruction manual 1

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 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
8. Add 50µL Stop Solution. Read at 450nm immediately.

ELISA Kit for Leptin (LEP)

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 Leptin (LEP). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to 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 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 Leptin (LEP) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Citations

  • Puerarin improves insulin resistance and modulates adipokine expression in rats fed a high-fat dietPubMed: 20869961
  • Effect of Argyreia speciosa root extract on cafeteria diet-induced obesity in ratsIjp: source
  • Testosterone protects from metabolic syndrome-associated prostate inflammation: an experimental study in rabbitPubMed: 22010203
  • Multi-Functional Roles of Chitosan as a Potential Protective Agent against ObesityPubMed: 23342013
  • Soy isoflavones modulate adipokines and myokines to regulate lipid metabolism in adipose tissue, skeletal muscle and liver of male Huanjiang mini-pigsScienceDirect: S0303720712004194
  • Ovarian, metabolic and endocrine indexes in dairy cows with different body condition scoresIfzz: Source
  • n-6: n-3 PUFA ratio is involved in regulating lipid metabolism and inflammation in pigsPubmed: 23947577
  • Maternal high folic acid supplement promotes glucose intolerance and insulin resistance in male mouse offspring fed a high-fat diet.Pubmed: 24736781
  • CTRP3 modulates the expression and secretion of adipokines in 3T3-L1 adipocytesPubmed:25168658
  • EFFECT OF LEPTIN GENE ON SERUM LEPTIN CONCENTRATION AND BEEF QUALITY IN CZECH FLECKVIEH BULLSMendelu:Source
  • Tinospora crispa Ameliorates Insulin Resistance Induced by High Fat Diet in Wistar RatsPubmed:Pmc4363645
  • Oxytocin reverse ovariectomy-induced Osteoper and body fat gainPubmed:24506069
  • Maternal high-fat diet feeding during pregnancy and lactation augments lung inflammation and remodeling in the offspringPubmed:25500158
  • Hepatic steatosis depresses alpha-1-antitrypsin levels in human and rat acute pancreatitisPubMed: 26634430
  • Tinospora crispaPubMed: 25821506
  • Effects of breeder age and early energy restriction on fattening performance, some meat quality traits and plasma leptin concentration of broilersDergiler: 11
  • Comparative measurements of the morphometric pattern and energy profile of Crioula mares in the final third of gestation, with different body condition scoresScielo: S0102-09352015000300707
  • Pancreatic Exocrine Insufficiency Affects Not Only Digestibility of Nutrients and Growth, but Also Body Composition and Endocrinological Parameters – Study on Piglets Used As a Model for Children Journals: 2165-7092-%20S5-00
  • Effect of extended photoperiod during winter on growth and onset of puberty in Murrah buffalo heifersPubmed:27051212
  • Anterior pituitary influence on adipokine expression and secretion by porcine adipocytes.Pubmed:27074715
  • Comparison of the effect of a standard inclusion level of inorganic zinc to organic form at lowered level on bone development in growing male Ross broiler chickens journalprintahead:87
  • Comparison of the Effect of a Standard Inclusion Level of Inorganic Zinc to Organic Form at Lowered Level on Bone Development in Growing Male Ross Broiler Chickensview:87
  • Changes on Adipose Tissue Distribution After Laparoscopic Roux-en-Y Gastric Bypass in Obese Göttingen Minipig. Effects on Glucose Metabolism.Pubmed:27185175
  • Favorable outcomes of hydroxychloroquine in insulin resistance may be accomplished by adjustment of the endothelial dysfunction as well as the skewed balance of adipokinesscience:S0065128116301234
  • Nanosized titanium dioxide resulted in the activation of TGF-β/Smads/p38MAPK pathway in renal inflammation and fibration of micePubmed:26850371
  • The bromodomain inhibitor N-methyl pyrrolidone reduced fat accumulation in an ovariectomized rat modelPubmed:27110299
  • Oral Supplementation with a Special Additive of Retinyl Palmitate and Alpha Tocopherol Reduces Growth Retardation in Young Pancreatic Duct Ligated Pigs Used as a Model for Children Suffering from Exocrine Pancreatic Insufficiencypubmed:27690005
  • Changes in small intestinal morphometry and electrogenic nutrient transport in young pigs with pancreatic exocrine insufficiencypublications:jas
  • The Changes of Serum Leptin and Kisspeptin Levels in Chinese Children and Adolescents in Different Pubertal Stagespubmed:27990162
  • The effect of GnRH vaccination on performance, carcass, and meat quality and hormonal regulation in boars, barrows, and gilts.pubmed:27482668
  • Diet-induced modifications to milk composition have long-term effects on offspring growth in rabbits.pubmed:28380600
  • Four-Week Consumption of Malaysian Honey Reduces Excess Weight Gainand Improves Obesity-Related Parameters in High Fat Diet Induced ObeseRats.pubmed:28246535
  • Dietary Niacin Supplementation Suppressed Hepatic Lipid Accumulation in Rabbitspubmed:27004817
  • Serial changes in the proliferation and differentiation of adipose-derived stem cells after ionizingradiation.pubmed:27530249
  • Changes on Adipose Tissue Distribution After Laparoscopic Roux-en-Y Gastric Bypass in ObeseGöttingen Minipig. Effects on Glucose Metabolism.pubmed:27185175
  • Estimation of the intramuscular fat content of m. longissimus thoracis in crossbred beef cattle based on live animal measurements.pubmed:27940421
  • Correlation of Leptin, Insulin and Glucose During Late Gestation and Early Lactation in Murrah Buffaloes7086
  • Effects Of Maca (Lepıdıum Meyenıı Walp) Powder On Serum Indıces And Metabolıc Responses In Racehorses57cf073008ae83b37462323a.pdf

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