Home / Leptin/OB / Mouse Leptin DuoSet ELISA

Mouse Leptin DuoSet ELISA

Catalog #: DY498
40 Citations 4 Reviews Datasheet / COA / SDS
Catalog # Availability Size / Price Qty
DY498
DY498-05
Ancillary Products Available
DY999B: Substrate Reagent Pack
DY999: Substrate Reagent Pack
DY009B: DuoSet ELISA Ancillary Reagent Kit 3
Mouse Leptin / OB ELISA Standard Curve
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Product Details
Procedure
Citations (40)
FAQs
Supplemental Products
Reviews (4)
Summary Product Features Kit Content Other Reagents Required Data Examples Product Datasheets Preparation and Storage Background

Mouse Leptin DuoSet ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Sample Volume Required
100 µL
Assay Range
125.0 - 8,000 pg/mL
Sufficient Materials
For fifteen 96-well plates*
Specificity
Please see the product datasheet

* Provided that the recommended microplates, buffers, diluents, substrates and solutions are used, and the assay is run as summarized in the Assay Procedure provided.

This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant mouse Leptin. The suggested diluent is suitable for the analysis of most cell culture supernate samples. Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet.

Product Features

  • Optimized capture and detection antibody pairings with recommended concentrations save lengthy development time
  • Development protocols are provided to guide further assay optimization
  • Assay can be customized to your specific needs
  • Economical alternative to complete kits

Kit Content

  • Capture Antibody
  • Detection Antibody
  • Recombinant Standard
  • Streptavidin conjugated to horseradish-peroxidase (Streptavidin-HRP)

Other Reagents Required

DuoSet Ancillary Reagent Kit 3 (5 plates): (Catalog # DY009) containing 96 well microplates, plate sealers, substrate solution, stop solution, plate coating buffer (PBS), wash buffer, and Reagent Diluent Concentrate 3.

The components listed above may be purchased separately:

PBS: (Catalog # DY006), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na2HPO4, 1.5 mM KH2PO4, pH 7.2 - 7.4, 0.2 µm filtered

Wash Buffer: (Catalog # WA126), or 0.05% Tween® 20 in PBS, pH 7.2-7.4

Reagent Diluent: (Catalog # DY004), or 5% Tween 20 in PBS, 7.2-7.4, 0.2 μm filtered

Substrate Solution: 1:1 mixture of Color Reagent A (H2O2) and Color Reagent B (Tetramethylbenzidine) (Catalog # DY999)

Stop Solution: 2 N H2SO4 (Catalog # DY994)

Microplates: R&D Systems (Catalog # DY990)

Plate Sealers: ELISA Plate Sealers (Catalog # DY992)

 

Scientific Data

N/A Mouse Leptin / OB ELISA Standard Curve View Larger

Mouse Leptin / OB ELISA Standard Curve

Product Datasheets

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Product Datasheet
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Preparation and Storage

Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: Leptin/OB

Leptin is a product of the mouse obese gene. Mice with mutations in the obese gene that block the synthesis of leptin are obese and diabetic, and have reduced activity, metabolism, and body temperature. Rat leptin shares approximately 96% and 82% sequence identity with the mouse and human protein, respectively. The expression of leptin mRNA is restricted to adipose tissue.

Entrez Gene IDs:
3952 (Human); 16846 (Mouse); 25608 (Rat)
Alternate Names:
FLJ94114; LEP; leptin (murine obesity homolog); leptin (obesity homolog, mouse); Leptin; OB; Obese protein; obese, mouse, homolog of; Obesity factor; OBOBS

Assay Procedure

GENERAL ELISA PROTOCOL


Plate Preparation

  1. Dilute the Capture Antibody to the working concentration in PBS without carrier protein. Immediately coat a 96-well microplate with 100 μL per well of the diluted Capture Antibody. Seal the plate and incubate overnight at room temperature.
  2. Aspirate each well and wash with Wash Buffer, repeating the process two times for a total of three washes. Wash by filling each well with Wash Buffer (400 μL) using a squirt bottle, manifold dispenser, or autowasher. Complete removal of liquid at each step is essential for good performance. After the last wash, remove any remaining Wash Buffer by aspirating or by inverting the plate and blotting it against clean paper towels.
  3. Block plates by adding 300 μL of Reagent Diluent to each well. Incubate at room temperature for a minimum of 1 hour.
  4. Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.

Assay Procedure

  1. Add 100 μL of sample or standards in Reagent Diluent, or an appropriate diluent, per well. Cover with an adhesive strip and incubate 2 hours at room temperature.
  2. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  3. Add 100 μL of the Detection Antibody, diluted in Reagent Diluent, to each well. Cover with a new adhesive strip and incubate 2 hours at room temperature.
  4. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  5. Add 100 μL of the working dilution of Streptavidin-HRP to each well. Cover the plate and incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  6. Repeat the aspiration/wash as in step 2.
  7. Add 100 μL of Substrate Solution to each well. Incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  8. Add 50 μL of Stop Solution to each well. Gently tap the plate to ensure thorough mixing.
  9. Determine the optical density of each well immediately, using a microplate reader set to 450 nm. If wavelength correction is available, set to 540 nm or 570 nm. If wavelength correction is not available, subtract readings at 540 nm or 570 nm from the readings at 450 nm. This subtraction will correct for optical imperfections in the plate. Readings made directly at 450 nm without correction may be higher and less accurate.

Citations for Mouse Leptin DuoSet ELISA

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

40 Citations: Showing 1 - 10
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  1. Diet switch pre-vaccination improves immune response and metabolic status in formerly obese mice
    Authors: Honce, R;Vazquez-Pagan, A;Livingston, B;Mandarano, AH;Wilander, BA;Cherry, S;Hargest, V;Sharp, B;Brigleb, PH;Kirkpatrick Roubidoux, E;Van de Velde, LA;Skinner, RC;McGargill, MA;Thomas, PG;Schultz-Cherry, S;
    Nature microbiology
    Species: Mouse
    Sample Types: Serum
  2. Thyme Extract Alleviates High-Fat Diet-Induced Obesity and Gut Dysfunction
    Authors: Lee, YR;Lee, HB;Oh, MJ;Kim, Y;Park, HY;
    Nutrients
    Species: Mouse
    Sample Types: Serum
  3. Chronic hyperpalatable diet induces impairment of hippocampal-dependent memories and alters glutamatergic and fractalkine axis signaling
    Authors: Ribeiro, R;Silva, EG;Moreira, FC;Gomes, GF;Cussat, GR;Silva, BSR;da Silva, MCM;de Barros Fernandes, H;de Sena Oliveira, C;de Oliveira Guarnieri, L;Lopes, V;Ferreira, CN;de Faria, AMC;Maioli, TU;Ribeiro, FM;de Miranda, AS;Moraes, GSP;de Oliveira, ACP;Vieira, LB;
    Scientific reports
    Species: Mouse
    Sample Types: Serum, Tissue Homogenates
  4. HILPDA is a lipotoxic marker in adipocytes that mediates the autocrine negative feedback regulation of triglyceride hydrolysis by fatty acids and alleviates cellular lipotoxic stress
    Authors: Deng, L;Wu, SA;Qi, L;Kersten, S;
    Molecular metabolism
    Species: Mouse
    Sample Types: Plasma
  5. mTORC2-NDRG1-CDC42 axis couples fasting to mitochondrial fission
    Authors: Martinez-Lopez, N;Mattar, P;Toledo, M;Bains, H;Kalyani, M;Aoun, ML;Sharma, M;McIntire, LBJ;Gunther-Cummins, L;Macaluso, FP;Aguilan, JT;Sidoli, S;Bourdenx, M;Singh, R;
    Nature cell biology
  6. Long-term statins administration exacerbates diabetic nephropathy via ectopic fat deposition in diabetic mice
    Authors: TS Huang, T Wu, YD Wu, XH Li, J Tan, CH Shen, SJ Xiong, ZQ Feng, SF Gao, H Li, WB Cai
    Nature Communications, 2023-01-24;14(1):390.
    Species: Mouse
    Sample Types: Serum
  7. Early macrophage response to obesity encompasses Interferon Regulatory Factor 5 regulated mitochondrial architecture remodelling
    Authors: L Orliaguet, T Ejlalmanes, A Humbert, R Ballaire, M Diedisheim, JB Julla, D Chokr, J Cuenco, J Michielett, J Charbit, D Lindén, J Boucher, C Potier, A Hamimi, S Lemoine, C Blugeon, P Legoix, S Lameiras, LG Baudrin, S Baulande, A Soprani, FA Castelli, F Fenaille, JP Riveline, E Dalmas, J Rieusset, JF Gautier, N Venteclef, F Alzaid
    Nature Communications, 2022-08-30;13(1):5089.
    Species: Mouse
    Sample Types: Plasma
  8. Characterization of autochthonous lactobacilli from goat dairy products with probiotic potential for metabolic diseases
    Authors: A Marquez, E Andrada, M Russo, ML Bolondi, E Fabersani, R Medina, P Gauffin-Ca
    Heliyon, 2022-08-28;8(9):e10462.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  9. Exogenous leptin enhances markers of airway fibrosis in a mouse model of chronic allergic airways disease
    Authors: MD Ihrie, VL McQuade, JT Womble, A Hegde, MS McCravy, CVG Lacuesta, RM Tighe, LG Que, JKL Walker, JL Ingram
    Respiratory Research, 2022-05-24;23(1):131.
    Species: Mouse
    Sample Types: Serum
  10. Impact of controlled high-sucrose and high-fat diets on eosinophil recruitment and cytokine content in allergen-challenged mice
    Authors: CM Percopo, M McCullough, AR Limkar, KM Druey, HF Rosenberg
    PLoS ONE, 2021-08-12;16(8):e0255997.
    Species: Mouse
    Sample Types: BALF
  11. The autophagy protein Becn1 improves insulin sensitivity by promoting adiponectin secretion via exocyst binding
    Authors: K Kuramoto, YJ Kim, JH Hong, C He
    Cell Reports, 2021-05-25;35(8):109184.
    Species: Mouse
    Sample Types: Serum
  12. C1q/TNF-Related Protein 3 (CTRP-3) Deficiency of Adipocytes Affects White Adipose Tissue Mass but Not Systemic CTRP-3 Concentrations
    Authors: A Schmid, M Roderfeld, J Gehl, E Roeb, A Nist, HR Chung, T Stiewe, T Karrasch, A Schäffler
    International Journal of Molecular Sciences, 2021-02-07;22(4):.
    Species: Mouse
    Sample Types: Serum
  13. Effect of Lactobacillus rhamnosus GG on Energy Metabolism, Leptin Resistance, and Gut Microbiota in Mice with Diet-Induced Obesity
    Authors: YC Cheng, JR Liu
    Nutrients, 2020-08-24;12(9):.
    Species: Mouse
    Sample Types: Serum
  14. HILPDA Uncouples Lipid Droplet Accumulation in Adipose Tissue Macrophages from Inflammation and Metabolic Dysregulation
    Authors: XAMH van Dieren, MA de la Rosa, A Georgiadi, F Mattijssen, W Dijk, M van Weeghe, R Singh, JW Borst, R Stienstra, S Kersten
    Cell Rep, 2020-02-11;30(6):1811-1822.e6.
    Species: Mouse
    Sample Types: Plasma
  15. CSF1R inhibitor PLX5622 and environmental enrichment additively improve metabolic outcomes in middle-aged female mice
    Authors: S Ali, AG Mansour, W Huang, NJ Queen, X Mo, JM Anderson, QN Hassan Ii, RS Patel, RK Wilkins, MA Caligiuri, L Cao
    Aging (Albany NY), 2020-02-02;12(0):.
    Species: Mouse
    Sample Types: Serum
  16. The role of uncoupling protein 2 in macrophages and its impact on obesity-induced adipose tissue inflammation and insulin resistance
    Authors: XAMH van Dieren, T Sancerni, MC Alves-Guer, R Stienstra
    J Biol Chem, 2020-01-01;0(0):.
    Species: Mouse
    Sample Types: Plasma
  17. SOCS2 modulates adipose tissue inflammation and expansion in mice
    Authors: CH Val, MC de Oliveir, DR Lacerda, A Barroso, NV Batista, Z Menezes-Ga, DRR de Assis, AT Cramer, F Brant, MM Teixeira, D Glória Sou, AM Ferreira, FS Machado
    J. Nutr. Biochem., 2019-11-23;76(0):108304.
    Species: Mouse
    Sample Types: Serum
  18. Environmental enrichment improves metabolic and behavioral health in the BTBR mouse model of autism
    Authors: NJ Queen, AA Boardman, RS Patel, JJ Siu, X Mo, L Cao
    Psychoneuroendocrinology, 2019-10-15;111(0):104476.
    Species: Mouse
    Sample Types: Serum
  19. Adipose PTEN regulates adult adipose tissue homeostasis and redistribution via a PTEN-leptin-sympathetic loop
    Authors: W Huang, NJ Queen, TB McMurphy, S Ali, L Cao
    Mol Metab, 2019-09-28;30(0):48-60.
    Species: Mouse
    Sample Types: Serum
  20. Melatonin Supplementation Attenuates the Pro-Inflammatory Adipokines Expression in Visceral Fat from Obese Mice Induced by A High-Fat Diet
    Authors: TDSM Farias, RID Paixao, MM Cruz, RDCDC de Sa, JJ Simão, VJ Antraco, MIC Alonso-Val
    Cells, 2019-09-06;8(9):.
    Species: Mouse
    Sample Types: Tissue Lysates
  21. Resistance to visceral obesity is associated with increased locomotion in mice expressing an endothelial cell-specific fibroblast growth factor 1 transgene
    Authors: T Keeley, A Kirov, WY Koh, V Demambro, I Bergquist, J Cotter, P Caradonna, ME Siviski, B Best, T Henderson, CJ Rosen, L Liaw, I Prudovsky, DJ Small
    Physiol Rep, 2019-04-01;7(7):e14034.
    Species: Mouse
    Sample Types: Serum
  22. GPS2 Deficiency Triggers Maladaptive White Adipose Tissue Expansion in Obesity via HIF1A Activation
    Authors: K Drareni, R Ballaire, S Barilla, MJ Mathew, A Toubal, R Fan, N Liang, C Chollet, Z Huang, M Kondili, F Foufelle, A Soprani, R Roussel, JF Gautier, F Alzaid, E Treuter, N Venteclef
    Cell Rep, 2018-09-11;24(11):2957-2971.e6.
    Species: Mouse
    Sample Types: Whole Blood
  23. Metformin Mitigates Fibrosis and Glucose Intolerance Induced by Doxorubicin in Subcutaneous Adipose Tissue
    Authors: LA Biondo, HA Batatinha, CO Souza, AAS Teixeira, LS Silveira, MI Alonso-Val, LM Oyama, MJ Alves, M Seelaender, JCR Neto
    Front Pharmacol, 2018-05-08;9(0):452.
    Species: Mouse
    Sample Types: Serum
  24. Collagen overlays can inhibit leptin and adiponectin secretion but not lipid accumulation in adipocytes
    Authors: SL Christian, NK Pallegar, RJ Brown, AM Viloria-Pe
    PeerJ, 2018-04-27;6(0):e4641.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  25. Unique metabolic activation of adipose tissue macrophages in obesity promotes inflammatory responses
    Authors: L Boutens, GJ Hooiveld, S Dhingra, RA Cramer, MG Netea, R Stienstra
    Diabetologia, 2018-01-14;0(0):.
    Species: Mouse
    Sample Types: Tissue Culture Supernates
  26. Inducing maternal inflammation promotes leptin production in offspring but does not improve allergic symptoms in a mouse model of allergic rhinitis
    Authors: A Imai, K Satoi, E Fujimoto, K Sato
    Heliyon, 2017-06-22;3(6):e00327.
    Species: Mouse
    Sample Types: Serum
  27. Combination of a high-fat diet with sweetened condensed milk exacerbates inflammation and insulin resistance induced by each separately in mice
    Authors: LN Masi, AR Martins, AR Crisma, CL do Amaral, MR Davanso, TDA Serdan, RDC da Cunha d, MM Cruz, MIC Alonso-Val, RP Torres, J Mancini-Fi, JNB Pereira, MM da Silva R, EA Liberti, SM Hirabara, R Curi
    Sci Rep, 2017-06-21;7(1):3937.
    Species: Mouse
    Sample Types: Tissue Homogenates
  28. Targeting Visceral Fat by Intraperitoneal Delivery of Novel AAV Serotype Vector Restricting Off-Target Transduction in Liver
    Authors: W Huang, X Liu, NJ Queen, L Cao
    Mol Ther Methods Clin Dev, 2017-06-19;6(0):68-78.
    Species: Mouse
    Sample Types: Serum
  29. Hepatic Expression of Adenovirus 36 E4ORF1 Improves Glycemic Control and Promotes Glucose Metabolism via AKT Activation
    Authors: Travis B McMurphy
    Diabetes, 2016-11-30;0(0):.
    Species: Mouse
    Sample Types: Serum
  30. Loss of CD24 in Mice Leads to Metabolic Dysfunctions and a Reduction in White Adipocyte Tissue.
    Authors: Fairbridge N, Southall T, Ayre D, Komatsu Y, Raquet P, Brown R, Randell E, Kovacs C, Christian S
    PLoS ONE, 2015-11-04;10(11):e0141966.
    Species: Mouse
    Sample Types: Plasma
  31. Obesity increases airway hyperresponsiveness via the TNF-alpha pathway and treating obesity induces recovery.
    Authors: Kim J, Sohn J, Lee J, Park J
    PLoS ONE, 2015-02-06;10(2):e0116540.
    Species: Mouse
    Sample Types: Tissue Homogenates
  32. Interleukin-15 modulates adipose tissue by altering mitochondrial mass and activity.
    Authors: Barra, Nicole G, Palanivel, Rengasam, Denou, Emmanuel, Chew, Marianne, Gillgrass, Amy, Walker, Tina D, Kong, Josh, Richards, Carl D, Jordana, Manel, Collins, Stephen, Trigatti, Bernardo, Holloway, Alison C, Raha, Sandeep, Steinberg, Gregory, Ashkar, Ali A
    PLoS ONE, 2014-12-17;9(12):e114799.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  33. Hypothalamic gene transfer of BDNF inhibits breast cancer progression and metastasis in middle age obese mice.
    Authors: Liu, Xianglan, McMurphy, Travis, Xiao, Run, Slater, Andrew, Huang, Wei, Cao, Lei
    Mol Ther, 2014-03-18;22(7):1275-84.
    Species: Mouse
    Sample Types: Serum
  34. Increased expression of c-Jun in nonalcoholic fatty liver disease.
    Authors: Dorn C, Engelmann J, Saugspier M, Koch A, Hartmann A, Muller M, Spang R, Bosserhoff A, Hellerbrand C
    Lab Invest, 2014-02-03;94(4):394-408.
    Species: Mouse
    Sample Types: Serum
  35. Lack of platelet-activating factor receptor protects mice against diet-induced adipose inflammation and insulin-resistance despite fat pad expansion.
    Authors: Menezes-Garcia Z, Oliveira M, Lima R, Soriani F, Cisalpino D, Botion L, Teixeira M, Souza D, Ferreira A
    Obesity (Silver Spring), 2013-12-14;22(3):663-72.
    Species: Mouse
    Sample Types: Serum
  36. A2B adenosine receptors prevent insulin resistance by inhibiting adipose tissue inflammation via maintaining alternative macrophage activation.
    Authors: Csoka B, Koscso B, Toro G, Kokai E, Virag L, Nemeth Z, Pacher P, Bai P, Hasko G
    Diabetes, 2013-11-05;63(3):850-66.
    Species: Mouse
    Sample Types: Plasma
  37. Mutant amyloid precursor protein differentially alters adipose biology under obesogenic and non-obesogenic conditions.
    Authors: Freeman, Linnea R, Zhang, Le, Dasuri, Kalavath, Fernandez-Kim, Sun-Ok, Bruce-Keller, Annadora, Keller, Jeffrey
    PLoS ONE, 2012-08-17;7(8):e43193.
    Species: Mouse
    Sample Types: Serum
  38. Rosiglitazone improves survival and hastens recovery from pancreatic inflammation in obese mice.
    Authors: Pini M, Rhodes DH, Castellanos KJ
    PLoS ONE, 2012-07-16;7(7):e40944.
    Species: Mouse
    Sample Types: Plasma
  39. Neuropeptide Y acts directly in the periphery on fat tissue and mediates stress-induced obesity and metabolic syndrome.
    Authors: Kuo LE, Kitlinska JB, Tilan JU, Li L, Baker SB, Johnson MD, Lee EW, Burnett MS, Fricke ST, Kvetnansky R, Herzog H, Zukowska Z
    Nat. Med., 2007-07-01;13(7):803-11.
    Species: Mouse
    Sample Types: Plasma
  40. Increased pulmonary responses to acute ozone exposure in obese db/db mice.
    Authors: Lu FL, Johnston RA, Flynt L, Theman TA, Terry RD, Schwartzman IN, Lee A, Shore SA
    Am. J. Physiol. Lung Cell Mol. Physiol., 2005-12-22;290(5):L856-65.
    Species: Mouse
    Sample Types: Serum

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Reviews for Mouse Leptin DuoSet ELISA

Average Rating: 5 (Based on 4 Reviews)

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Mouse Leptin DuoSet ELISA
By Carmen Martin-Ruiz on 02/07/2020
Sample Tested: Mouse serum and protein extract

Very straight forward, we managed to bring the suggested standard curve down to 62.5 pg/ml. Very reliable. Serum diluted 1:5, protein homogenates diluted 1:2

Mouse Leptin DuoSet ELISA DY498
Mouse Leptin DuoSet ELISA
By Ivan Luzardo on 12/11/2018
Sample Tested: Serum and Plasma
Mouse Leptin DuoSet ELISA
By Anonymous on 05/04/2018
Sample Tested: Liver cancer
Mouse Leptin DuoSet ELISA
By Anonymous on 04/11/2018
Sample Tested: Pancreatic cancer cells