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|Antigen||Leptin (LEP) ELISA Kits|
|Reactivity||Pig (Porcine), Wild boar (Sus scrofa) Alternatives|
Kits with alternative reactivity to:
|Method Type||Sandwich ELISA|
|Detection Range||31.2 pg/mL - 2000 pg/mL|
|Minimum Detection Limit||31.2 pg/mL|
|8 references available|
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Product Details Leptin ELISA KitTarget Details Application Details Handling References for Leptin Kit (ABIN415853) Images
|Purpose||The kit is a sandwich enzyme immunoassay for in vitro quantitative measurement of LEP in Serum,Plasma,Tissue Homogenate,Biological Fluids|
|Sample Type||Plasma, Serum, Tissue Homogenate|
This assay has high sensitivity and excellent specificity for detection of Leptin (LEP).
|Cross-Reactivity (Details)||No significant cross-reactivity or interference between Leptin (LEP) and analogues was observed.|
|Material not included||
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|Alternative Name||LEP (LEP ELISA Kit Abstract)|
|Pathways||JAK-STAT Signaling, AMPK Signaling, Hormone Transport, Peptide Hormone Metabolism, Hormone Activity, Negative Regulation of Hormone Secretion, Regulation of Carbohydrate Metabolic Process, Feeding Behaviour, Monocarboxylic Acid Catabolic Process|
Application DetailsProduct Details Leptin ELISA Kit Target Details Handling References for Leptin Kit (ABIN415853) Images back to top
Information on standard material:
|Sample Volume||100 μL|
|Assay Time||3 h|
|Protocol||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.|
Serum: Allow samples to clot for two hours at room temperature or overnight at 4°C before centrifugation for 20 minutes at approximately 1000 × g. Assay immediately or store samples in aliquot at -20°C or -80°C. Avoid repeated freeze/thaw cycles.
Plasma: Collect plasma using EDTA or heparin as an anticoagulant. Centrifuge samples for 15 minutes at 1000 × g within 30 minutes of collection. Remove plasma and assay immediately or store samples in aliquot at -20°C or -80°C. Avoid repeated freeze/thaw cycles.
Tissue Homogenates: The preparation of tissue homogenates will vary depending upon tissue type. For this assay, rinse tissues in ice-cold PBS (0.02mol/L,pH 7.0-7.2) to remove excess blood thoroughly and weigh before homogenization. Mince the tissues to small pieces and homogenize them in 5-10 mL of PBS with a glass homogenizer on ice (Micro Tissue Grinders work, too). Sonicate the resulting suspension with an ultrasonic cell disrupter or subject it to two freeze-thaw cycles to further break the cell membranes. Centrifugate the homogenates for 5 minutes at 5000 × g. Remove the supernate and assay immediately or aliquot and store at -20°C
Biological Fluids: Centrifuge samples for 20 minutes at 1000 × g. Remove particulates and assay immediately or store samples in aliquot at -20 °C or -80 °C for later use. Avoid repeated freeze/thaw cycles.
|Calculation of Results||
Average the duplicate readings for each standard, control, and samples and subtract the average zero standard optical density. Construct a standard curve by plotting the mean O.D. and concentration for each standard and draw a best fit curve through the points on the graph or create a standard curve on log-log graph paper with LEP concentration on the y-axis and absorbance on the x-axis. Using some plot software, for instance, curve expert 1.30, is also recommended. If samples have been diluted, the concentration read from the standard curve must be multiplied by the dilution factor.
In order to make the calculation easier, we plot the O.D. value of the standard (X-axis) against the known concentration of the standard (Y-axis), although concentration is the independent variable and O.D. value is the dependent variable. However, the O.D. values of the standard curve may vary according to the conditions of assay performance (e.g. operator, pipetting technique, washing technique or temperature effects), plotting log of the data to establish standard curve for each test is recommended. Typical standard curve below is provided for reference only.
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.
|Restrictions||For Research Use only|
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|Precaution of Use||The Stop Solution suggested for use with this kit is an acid solution. Wear eye, hand, face, and clothing protection when using this material.|
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.
|Expiry Date||6 months|
References for Leptin Kit (ABIN415853)Product Details Leptin ELISA Kit Target Details Application Details Handling Images back to top
|Product cited in:||
Hernández Hurtado, Sánchez-Margallo, De la Cruz Vigo, Maestre Antequera, Matos Azevedo, Casado, Díaz-Güemes Martín-Portugués: "Changes on Adipose Tissue Distribution After Laparoscopic Roux-en-Y Gastric Bypass in Obese Göttingen Minipig. Effects on Glucose Metabolism." in: Obesity surgery, 2016
Saleri, Cavalli, Martelli, Borghetti: "Anterior pituitary influence on adipokine expression and secretion by porcine adipocytes." in: Animal : an international journal of animal bioscience, Vol. 10, Issue 6, pp. 933-8, 2016
Van den Broeke, Leen, Aluwé, Ampe, Van Meensel, Millet: "The effect of GnRH vaccination on performance, carcass, and meat quality and hormonal regulation in boars, barrows, and gilts." in: Journal of animal science, Vol. 94, Issue 7, pp. 2811-20, 2016
Jeong, Yang, Lee, Ryoo, Kim, Oh, Kwon, Liu, Son: "Serial changes in the proliferation and differentiation of adipose-derived stem cells after ionizing radiation." in: Stem cell research & therapy, Vol. 7, Issue 1, pp. 117, 2016
Mößeler, Schmicke, Höltershinken, Beyerbach, Kamphues et al.: "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 ..." in: International journal of molecular sciences, Vol. 17, Issue 10, 2016
Duan, Li, Li, Fan, Sun, Yin: "n-6:n-3 PUFA ratio is involved in regulating lipid metabolism and inflammation in pigs." in: The British journal of nutrition, Vol. 111, Issue 3, pp. 445-51, 2014
Walsh, Sweeney, Bahar, ODoherty: "Multi-functional roles of chitosan as a potential protective agent against obesity." in: PLoS ONE, Vol. 8, Issue 1, pp. e53828, 2013
Yang, Li, Xiong, Kong, Zhang, Yuan, Fan, Duan, Geng, Li, Yin: "Soy isoflavones modulate adipokines and myokines to regulate lipid metabolism in adipose tissue, skeletal muscle and liver of male Huanjiang mini-pigs." in: Molecular and cellular endocrinology, Vol. 365, Issue 1, pp. 44-51, 2012
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