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|Antigen||Leptin (LEP) ELISA Kits|
|Reactivity||Rat (Rattus) Alternatives|
Kits with alternative reactivity to:
|Method Type||Sandwich ELISA|
|Detection Range||30-8000 pg/mL|
|Minimum Detection Limit||30 pg/mL|
|10 references available|
|Supplier||Log in to see|
Product Details Leptin ELISA KitTarget details Application Details Handling References for Leptin Kit (ABIN625206) Images
|Purpose||Rat Leptin ELISA Kit for cell culture supernatants, plasma, and serum samples.|
|Sample Type||Plasma, Cell Culture Supernatant, Serum|
|Specificity||The Leptin ELISA kit shows no cross-reactivity with the following cytokines tested: rat CINC-2, CINC-3, CNTF, Fractalkine, IL-1 alpha, IL-1 beta, IL-4, IL-6, IL-10, GM-CSF, IFN-gamma, L-Selectin, Lix, MCP-1, MIP-3 alpha, beta-NGF, TIMP-1, TNF-alpha.|
|Cross-Reactivity (Details)||This ELISA kit shows no cross-reactivity with the following cytokines tested: rat CINC-2, CINC-3, CNTF, Fractalkine, IL-1alpha, IL-1beta, IL-4, IL-6, IL-10, GM-CSF, IFN-gamma, L-Selectin, Lix, MCP-1, MIP-3alpha, beta-NGF, TIMP-1, TNF-alpha.|
|Sensitivity||< 30 pg/mL|
|Material not included||
Target detailsProduct Details Leptin ELISA Kit Application Details Handling References for Leptin Kit (ABIN625206) Images back to top
|Alternative Name||Leptin (LEP ELISA Kit Abstract)|
|Background||Lens epithelial cell protein LEP503|
|Research Area||Cardiovascular, Atherosclerosis, Hormones|
|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 (ABIN625206) Images back to top
|Application Notes||Recommended Dilution for serum and plasma samples3 fold|
|Sample Volume||100 μL|
1. Bring all reagents and samples to room temperature (18 - 25 °C) before use.
2. Sample dilution: If your samples need to be diluted, 1x Assay Diluent D (Item K) should be used for dilution of serum/plasma/ culture supernatants. Suggested dilution for normal serum/plasma: 3 fold*. *Please note that levels of the target protein may vary between different specimens. Optimal dilution factors for each sample must be determined by the investigator.
3. Assay Diluent D (Item K) and Assay Diluent B (Item E) should be diluted 5-fold with deionized or distilled water before use.
4. Preparation of standard: Briefly spin the vial of Item C. Add 400 µL 1x Assay Diluent D (Item K) into Item C vial to prepare a 50 ng/mL standard solution. Dissolve the powder thoroughly by a gentle mix. Add 80 µL Leptin standard from the vial of Item C, into a tube with 420 µL 1x Assay Diluent D to prepare a 8,000 pg/mL standard solution. Pipette 400myl 1x Assay Diluent D into each tube. Use the stock standard solution to produce a dilution series . Mix each tube thoroughly before the next transfer. 1x Assay Diluent D serves as the zero standard (0 pg/mL). 200 µL 200 µL 200 µL 200 µL 200 µL 200myl 80 µL standard + 420 µL 8,000 2667 888.9 296.3 98.77 32.92 10.97 0 pg/mL pg/mL pg/mL pg/mL pg/mL pg/mL pg/mL pg/mL
5. If the Wash Concentrate (20x) (Item B) contains visible crystals, warm to room temperature and mix gently until dissolved. Dilute 20 ml of Wash Buffer Concentrate into deionized or distilled water to yield 400 ml of 1x Wash Buffer.
6. Briefly spin the Detection Antibody vial (Item F) before use. Add 100 µL of 1x Assay Diluent B (Item E) into the vial to prepare a detection antibody concentrate. Pipette up and down to mix gently (the concentrate can be stored at 4 °C for 5 days). The detection antibody concentrate should be diluted 80-fold with 1x Assay Diluent B and used in step 4 of Part VI Assay Procedure.
7. Briefly spin the HRP-Streptavidin concentrate vial (Item G) and pipette up and down to mix gently before use. HRP-Streptavidin concentrate should be diluted 200-fold with 1x Assay Diluent B (Item E). For example: Briefly spin the vial (Item G) and pipette up and down to mix gently . Add 50 µL of HRP-Streptavidin concentrate into a tube with 10 ml 1x Assay Diluent B to prepare a 200-fold diluted HRP-Streptavidin solution (don't store the diluted solution for next day use). Mix well.
1. Bring all reagents and samples to room temperature (18 - 25 °C) before use. It is recommended that all standards and samples be run at least in duplicate.
2. Add 100 µL of each standard (see Reagent Preparation step 2) and sample into appropriate wells. Cover well and incubate for 2.5 hours at room temperature or over night at 4 °C with gentle shaking.
3. Discard the solution and wash 4 times with 1x Wash Solution. Wash by filling each well with Wash Buffer (300 myl) using a multi-channel Pipette or autowasher. Complete removal of liquid at each step is essential to good performance. After the last wash, remove any remaining Wash Buffer by aspirating or decanting. Invert the plate and blot it against clean paper towels.
4. Add 100 µL of 1x prepared biotinylated antibody (Reagent Preparation step 6) to each well. Incubate for 1 hour at room temperature with gentle shaking.
5. Discard the solution. Repeat the wash as in step
6. Add 100 µL of prepared Streptavidin solution (see Reagent Preparation step 7) to each well. Incubate for 45 minutes at room temperature with gentle shaking.
7. Discard the solution. Repeat the wash as in step
8. Add 100 µL of TMB One-Step Substrate Reagent (Item H) to each well. Incubate for 30 minutes at room temperature in the dark with gentle shaking.
9. Add 50 µL of Stop Solution (Item I) to each well. Read at 450 nm immediately.
|Calculation of Results||
Calculate the mean absorbance for each set of duplicate standards, controls and samples, and subtract the average zero standard optical density. Plot the standard curve on log-log graph paper or using Sigma plot software, with standard concentration on the x-axis and absorbance on the y-axis. Draw the best-fit straight line through the standard points.
Typical Data: These standard curves are for demonstration only. A standard curve must be run with each assay. Rat Leptin concentration (pg/mL) O D =4 50 n m 0.01 0.1 1 10 Assay Diluent D 10 100 1,000 10,000
Sensitivity: The minimum detectable dose of Leptin is typically less than 30 pg/mL.
Recovery: Recovery was determined by spiking various levels of Leptin into normal rat serum, plasma and cell culture media. Mean recoveries are as follows: Sample Type Average % Recovery Range ( %) Serum 92.32 83-102 Plasma 84.25 75-94 Cell culture media 75.43 68-89
Linearity: Sample Type Serum Plasma Cell Culture Media 1:2 Average % of Expected 105.3 111.6 112.6 Range ( %) 94-113 102-122 103-124 1:4 Average % of Expected 108.1 119.2 113.5 Range ( %) 95-114 107-125 103-125
Reproducibility: Intra-Assay: CV<10 % Inter-Assay: CV<12 %
|Assay Precision||Intra-Assay: CV< 10 % Inter-Assay: CV< 12 %|
|Restrictions||For Research Use only|
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|Handling Advice||Avoid repeated freeze-thaw cycles.|
|Storage Comment||The entire kit may be stored at -20°C for up to 1 year from the date of shipment. Avoid repeated freeze-thaw cycles. The kit may be stored at 4°C for up to 6 months. For extended storage, it is recommended to store at -80°C.|
|Expiry Date||6 months|
References for Leptin Kit (ABIN625206)Product Details Leptin ELISA Kit Target details Application Details Handling Images back to top
|Product cited in:||
Govic, Penman, Tammer, Paolini: "Paternal calorie restriction prior to conception alters anxiety-like behavior of the adult rat progeny." in: Psychoneuroendocrinology, Vol. 64, pp. 1-11, 2016
Gooda Sahib Jambocus, Saari, Ismail, Khatib, Mahomoodally, Abdul Hamid: "An Investigation into the Antiobesity Effects of Morinda citrifolia L. Leaf Extract in High Fat Diet Induced Obese Rats Using a (1)H NMR Metabolomics Approach." in: Journal of diabetes research, Vol. 2016, pp. 2391592, 2016
Tawfik, Mahmoud, Saad, Shehata, Kamel, Helmy: "Similar and additive effects of ovariectomy and diabetes on insulin resistance and lipid metabolism." in: Biochemistry research international, Vol. 2015, pp. 567945, 2015
de Castro, Deminice, Simões-Ambrosio, Calder, Jordão, Vannucchi: "Dietary docosahexaenoic acid and eicosapentaenoic acid influence liver triacylglycerol and insulin resistance in rats fed a high-fructose diet." in: Marine drugs, Vol. 13, Issue 4, pp. 1864-81, 2015
Saad, Kamel, Hanafi: "Modulation of Adipocytokines Production and Serum NEFA Level by Metformin, Glimepiride, and Sitagliptin in HFD/STZ Diabetic Rats." in: Biochemistry research international, Vol. 2015, pp. 138134, 2015
Seif El-Din, El-Lakkany, El-Naggar, Hammam, Abd El-Latif, Ain-Shoka, Ebeid: "Effects of rosuvastatin and/or β-carotene on non-alcoholic fatty liver in rats." in: Research in pharmaceutical sciences, Vol. 10, Issue 4, pp. 275-87, 2015
Zhang, Wang, Long, Wang, Li, Gao, Tian: "Alteration of sweet taste in high-fat diet induced obese rats after 4 weeks treatment with exenatide." in: Peptides, Vol. 47, pp. 115-23, 2013
Huang, Korivi, Tsai, Yang, Tsai: "Supplementation of Lactobacillus plantarum K68 and Fruit-Vegetable Ferment along with High Fat-Fructose Diet Attenuates Metabolic Syndrome in Rats with Insulin Resistance." in: Evidence-based complementary and alternative medicine : eCAM, Vol. 2013, pp. 943020, 2013
Sloboda, Fève, Thornton, Nzietchueng, Regnault, Simon, Labat, Louis, Max, Muscat, Osborne-Pellegrin, Lacolley, Benetos: "Fatty acids impair endothelium-dependent vasorelaxation: a link between obesity and arterial stiffness in very old Zucker rats." in: The journals of gerontology. Series A, Biological sciences and medical sciences, Vol. 67, Issue 9, pp. 927-38, 2012
Leprat: "[Paraneoplastic endocrine syndromes]." in: La Revue du praticien, Vol. 15, Issue 15, pp. 2021-8, 1970
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