Mouse Leptin ELISA

RayBio® Mouse Leptin ELISA Kit for cell culture supernatants, plasma, and serum samples.

ELM-Leptin-1
+

Antigen Information

Accession Number:
  • P41160
Gene ID:
  • 16846
Gene Symbols:
  • LEP
  • OB
Protein Name & Synonyms:
Leptin (Obesity factor)
Target Species:
  • Mouse

Assay Format

Detection Range:
4 pg/ml - 1000 pg/ml
Recommended Dilution (serum & plasma):
10 - 40 fold
Sensitivity:
4 pg/ml

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Specificity:
This ELISA kit shows no cross-reactivity with any of the cytokines tested: Mouse CD30, L CD30, T CD40, CRG-2, CTACK, CXCL16, Eotaxin , Eotaxin-2, Fas Ligand, Fractalkine, GCSF, GM-CFS, IGFBP-3, IGFBP-5, IGFBP-6, IL-1 alpha, IL-1 beta, IL-2, IL-3, IL-3 Rb, IL-4, IL-5, IL-6, IL-9, IL-10, IL-12 p40/p70, IL-12 p70, IL-13, IL-17, KC, Leptin R, LIX, L-Selectin, Lymphotactin, MCP-1, MCP-5, M-CSF, MIG, MIP-1 alpha, MIP-1 gamma, MIP-2, MIP-3 beta, MIP-3 alpha, PF-4, P-Selectin, RANTES, SCF, SDF- 1alpha, TARC, TCA-3, TECK, TIMP-1, TNF-alpha, TNF RI, TNF RII, TPO, VCAM-1, VEGF.
Number of Targets Detected:
1
Species Detected:
  • Mouse
Compatible Sample Types:
  • Cell Culture Supernatants
  • Plasma
  • Serum
Design Principle:
  • Sandwich-based
Method of Detection:
Colorimetric
Quantitative/Semi-Quantitative:
Quantitative
Solid Support:
96-well Microplate

Product Specifications

Size:
1, 2, or 5 x 96-Well Strip Microplate Kit

Product Features

Additional Notes
  • Strip plates and additional reagents allow for use in multiple experiments
  • Quantitative protein detection
  • Establishes normal range
  • The best products for confirmation of antibody array data

Standard Curves

Spiking & Recovery Results

Linearity Results

Application Notes

Kit Components
  • Pre-Coated 96-well Strip Microplate
  • Wash Buffer
  • Stop Solution
  • Assay Diluent(s)
  • Lyophilized Standard
  • Biotinylated Detection Antibody
  • Streptavidin-Conjugated HRP
  • TMB One-Step Substrate
Other Materials Required
  • Distilled or deionized water
  • Precision pipettes to deliver 2 µl to 1 µl volumes
  • Adjustable 1-25 µl pipettes for reagent preparation
  • 100 µl and 1 liter graduated cylinders
  • Tubes to prepare standard and sample dilutions
  • Absorbent paper
  • Microplate reader capable of measuring absorbance at 450nm
  • Log-log graph paper or computer and software for ELISA data analysis
Protocol Outline
  1. Prepare all reagents, samples and standards as instructed in the manual.
  2. Add 100 µl of standard or sample to each well.
  3. Incubate 2.5 h at RT or O/N at 4°C.
  4. Add 100 µl of prepared biotin antibody to each well.
  5. Incubate 1 h at RT.
  6. Add 100 µl of prepared Streptavidin solution to each well.
  7. Incubate 45 min at RT.
  8. Add 100 µl of TMB One-Step Substrate Reagent to each well.
  9. Incubate 30 min at RT.
  10. Add 50 µl of Stop Solution to each well.
  11. Read at 450 nm immediately.

Storage/Stability

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.
  1. Eo H., Jeon Yj., Lee M., et al. Brown Alga Ecklonia cava Polyphenol Extract Ameliorates Hepatic Lipogenesis, Oxidative Stress, and Inflammation by Activation of AMPK and SIRT1 in High-Fat Diet-Induced Obese Mice. J Agric Food Chem. 2014 Dec 22.
    Species: Mouse
    Sample type: Plasma
  2. Park J., Shim H., Na A., et al. Orexin A regulates plasma insulin and leptin levels in a time-dependent manner following a glucose load in mice. Diabetologia. 2015 Jul;58(7):1542-50. doi: 10.1007/s00125-015-3573-0.
    Species: Mouse
    Sample type: Plasma
  3. Qian S-W, Tang Y, Li x, et al. BMP4-mediated brown fat-like changes in white adipose tissue alter glucose and energy homeostasis. Proc Natl Acad Sci USA. 2013 Feb 26;110(9):E798-807.
    Species: Mouse
    Sample type: Serum
  4. Wang X-A, Zhang R, Jiang D, et al. Interferon regulatory factor 9 protects against hepatic insulin resistance and steatosis in male mice. Hepatology. 2013 Aug;58(2):603-616.
    Species: Mouse
    Sample type: Serum
  5. Xia, Jia Nan, De Qin Zhang, Juan Du, and Jing Wen. "Regulation Effects of TZQ-F on Adipocyte Differentiation and Insulin Action." Journal of Ethnopharmacology. ScienceDirect, 25 Nov. 2013. Web.
    Species: Mouse
    Sample type: Serum
  6. Wang, Xin-An, et al. "CARD3 deficiency exacerbates diet-induced obesity, hepatosteatosis, and insulin resistance in male mice." Endocrinology 154.2 (2013): 685-697.
    Species: Mouse
    Sample type: Serum
  7. Wang, Xin-An, et al. "Interferon regulatory factor 7 deficiency prevents diet-induced obesity and insulin resistance." American Journal of Physiology-Endocrinology and Metabolism 305.4 (2013): E485-E495.
    Species: Mouse
    Sample type: Serum
  8. Naville, Danielle, et al. "Low-dose food contaminants trigger sex-specific, hepatic metabolic changes in the progeny of obese mice." The FASEB Journal 27.9 (2013): 3860-3870.
    Species: Mouse
    Sample type: Plasma
  9. Martín R, Hernández M, Córdova C, Nieto M. Natural triterpenes modulate immune-inflammatory markers of experimental autoimmune encephalomyelitis: therapeutic implications for multiple sclerosis. British Journal of Pharmacology 2012;166(5):1708-1723. doi:10.1111/j.1476-5381.2012.01869.x.
    Species: Mouse
    Sample type: Tissue Lysate
  10. Uner A., Sulu N. In vivo effects of leptin on lymphocyte subpopulations in mice. Immunobiology. 2012 Sep;217(9):882-8. doi: 10.1016/j.imbio.2011.12.013.
    Species: Mouse
    Sample type: Serum

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Usage

This product is furnished for LABORATORY RESEARCH USE ONLY.
Not for diagnostic or therapeutic use.