Mini Samples ELISA Kit for Leptin (LEP)

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

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.

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 87-101 98
EDTA plasma(n=5) 92-105 96
heparin plasma(n=5) 80-96 92

Precision

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

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Mini Samples 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) 98-105% 93-101% 79-97% 78-95%
EDTA plasma(n=5) 80-103% 80-104% 89-103% 97-104%
heparin plasma(n=5) 80-105% 78-95% 79-93% 88-95%

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

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 1×60µL Assay Diluent A 1×6mL
Detection Reagent B 1×60µL Assay Diluent B 1×6mL
TMB Substrate 1×4.5mL Stop Solution 1×3mL
Wash Buffer (30 × concentrate) 1×10mL Instruction manual 1

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.

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Magazine Citations
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Indian Journal of Pharmacology Effect of Argyreia speciosa root extract on cafeteria diet-induced obesity in rats Ijp: source
Journal of endocrinology Testosterone protects from metabolic syndrome-associated prostate inflammation: an experimental study in rabbit PubMed: 22010203
PLoS One. Multi-Functional Roles of Chitosan as a Potential Protective Agent against Obesity PubMed: 23342013
Molecular and Cellular Endocrinology Soy isoflavones modulate adipokines and myokines to regulate lipid metabolism in adipose tissue, skeletal muscle and liver of male Huanjiang mini-pigs ScienceDirect: S0303720712004194
Journal of Animal and Feed Sciences Ovarian, metabolic and endocrine indexes in dairy cows with different body condition scores Ifzz: Source
British Journal of Nutrition n-6: n-3 PUFA ratio is involved in regulating lipid metabolism and inflammation in pigs Pubmed: 23947577
Int J Mol Sci. Maternal high folic acid supplement promotes glucose intolerance and insulin resistance in male mouse offspring fed a high-fat diet. Pubmed: 24736781
Endocr J. CTRP3 modulates the expression and secretion of adipokines in 3T3-L1 adipocytes Pubmed:25168658
Animal Physiology EFFECT OF LEPTIN GENE ON SERUM LEPTIN CONCENTRATION AND BEEF QUALITY IN CZECH FLECKVIEH BULLS Mendelu:Source
Evid Based Complement Alternat Med. Tinospora crispa Ameliorates Insulin Resistance Induced by High Fat Diet in Wistar Rats Pubmed:Pmc4363645
general endocrinology Oxytocin reverse ovariectomy-induced Osteoper and body fat gain Pubmed:24506069
Respiratory Physiology &amp; Neurobiology Maternal high-fat diet feeding during pregnancy and lactation augments lung inflammation and remodeling in the offspring Pubmed:25500158
Sci Rep Hepatic steatosis depresses alpha-1-antitrypsin levels in human and rat acute pancreatitis PubMed: 26634430
Evid Based Complement Alternat Med Tinospora crispa PubMed: 25821506
Ankara Effects of breeder age and early energy restriction on fattening performance, some meat quality traits and plasma leptin concentration of broilers Dergiler: 11
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ANNALS OF ANIMAL SCIENCE 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
Annals of Animal Science 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 view:87
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journal of animal science The effect of GnRH vaccination on performance, carcass, and meat quality and hormonal regulation in boars, barrows, and gilts. pubmed:27482668
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Asian-Australasian Journal of Animal Sciences Dietary Niacin Supplementation Suppressed Hepatic Lipid Accumulation in Rabbits pubmed:27004817
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Oxidative Medicine and Cellular Longevity Changes of Plasma FABP4, CRP, Leptin, and Chemerin Levels in relation to Different Dietary Patterns and Duodenal-Jejunal Omega Switch Surgery in Sprague … Pubmed:29849871
Frontiers in Endocrinology Circulating and Adipose Tissue mRNA Levels of Zinc-α2-Glycoprotein, Leptin, High-Molecular-Weight Adiponectin, and Tumor Necrosis Factor-Alpha in … Pubmed:29755407
Journal of Agricultural and Food Chemistry Metabolomics for the effect of biotin and nicotinamide on transition dairy cows Pubmed:29758980
Frontiers in Physiology Ileal Transposition Surgery Decreases Fat Mass and Improves Glucose Metabolism in Diabetic GK Rats: Possible Involvement of FGF21 Pubmed:29593555
Bioscience Reports Effect of maternal hypothyroidism during pregnancy on insulin resistance, lipid accumulation and mitochondrial dysfunction in skeletal muscle of fetal rats Pubmed:29784871
Neurochemical Research The leptin, dopamine and serotonin receptors in hypothalamic POMC-neurons of normal and obese rodents Pubmed:29397535
PLoS One Subsequent somatic axis and bone tissue metabolism responses to a low-zinc diet with or without phytase inclusion in broiler chickens Pubmed:29373588
Biochemical and Biophysical Research Communications Chronic cold exposure results in subcutaneous adipose tissue browning and altered global metabolism in Qinghai-Tibetan plateau pika (Ochotona curzoniae) Pubmed:29626477
Alcohol and Alcoholism Leptin Expression and Gene Methylation Patterns in Alcohol-Dependent Patients with Ethyltoxic Cirrhosis—Normalization After Liver Transplantation and Implications … Pubmed:29912265
Iranian Red Crescent Medical Journal  The Effect of Aerobic Training on Tumor Growth, Adiponectin, Leptin and Ghrelin in Mice with Breast Cancer ircmj-20-03-13305.pdf
Pakistan Journal of Zoology Effects of Intrauterine Growth Retardation on Growth, Meat Quality and Muscle Fiber Composition of Pigs. :
Oncotarget Epigallocatechin-3-gallate suppresses differentiation of adipocytes via regulating the phosphorylation of FOXO1 mediated by PI3K-AKT signaling in 3T3-L1 … Pubmed:29484120
УСПЕХИ ГЕРОНТОЛОГИИ ВЛИЯНИЕ МЕТФОРМИНА НА МЕТАБОЛИЧЕСКИЕ ПОКАЗАТЕЛИ И ГИПОТАЛАМИЧЕСКИЕ СИГНАЛЬНЫЕ СИСТЕМЫ У КРЫС С ОЖИРЕНИЕМ … AG_2018-31-01.pdf
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