Leptin, also known as OB, is a polypeptide hormone that regulates adipose tissue mass and energy balance. In mice (ob/ob) and humans, inactivating mutations of the OB gene can cause obesity. Leptin signals through the transmembrane Leptin R in the hypothalamus but can also bind shorter and soluble forms of Leptin R which do not signal. Adipocytes increase Leptin expression as cell size increases, resulting in depressed appetite and increased energy expenditure. Low Leptin levels prompt increased appetite and decreased energy expenditure. Leptin can also block the onset of puberty or menses during starvation, extreme exercise-induced weight loss, anorexia, or cancer-induced cachexia.
Mouse/Rat Leptin Quantikine ELISA Kit
R&D Systems | Catalog # MOB00
Key Product Details
Assay Length
4.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (50 µL), Serum (20 µL), EDTA Plasma (20 µL)
Sensitivity
22 pg/mL
Assay Range
62.5-4000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma)
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Product Summary for Mouse/Rat Leptin Quantikine ELISA Kit
The Quantikine Mouse Leptin Immunoassay is a 4.5 hour solid phase ELISA designed to measure soluble mouse leptin in cell culture supernates, serum*, and plasma. It contains recombinant mouse leptin expressed in E. coli and antibodies raised against the recombinant factor. This immunoassay has been shown to quantitate the recombinant mouse leptin accurately. Results obtained measuring natural mouse leptin showed dose-response curves that were parallel to the standard curves obtained using the Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for natural mouse leptin.
*Pregnant mouse serum from the second trimester of pregnancy through post-partum day 1 require acid/urea activation of mouse leptin.
*Pregnant mouse serum from the second trimester of pregnancy through post-partum day 1 require acid/urea activation of mouse leptin.
Product Specifications
Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Measurement
Quantitative ELISA
Detection Method
Colorimetric - 450nm (TMB)
Conjugate
HRP
Species
Mouse, Rat
Specificity
Natural and recombinant mouse and rat Leptin. This assay cross-reacts 0.24% with Human Leptin.
Cross-reactivity
< 0.5% cross-reactivity observed with available related molecules. Cross-species reactivity observed with 1 or more species tested.
Interference
Interference observed with 1 or more available related molecules.
Precision
Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision.
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in separate assays to assess inter-assay precision.
Cell Culture Supernates, EDTA Plasma, Serum
| Intra-Assay Precision | Inter-Assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean (pg/mL) | 182 | 693 | 1803 | 187 | 654 | 1736 |
| Standard Deviation | 7.9 | 22.6 | 67.9 | 14.2 | 30.5 | 87.4 |
| CV% | 4.3 | 3.3 | 3.8 | 7.6 | 4.7 | 5.0 |
Recovery for Mouse/Rat Leptin Quantikine ELISA Kit
The recovery of mouse/rat Leptin spiked to three levels throughout the range of the assay in various matrices was evaluated.
| Sample Type | Average % Recovery | Range % |
|---|---|---|
| Mouse Cell Culture Supernates (n=5) | 101 | 85-120 |
| Mouse EDTA Plasma (n=4) | 92 | 78-105 |
| Mouse Heparin Plasma (n=4) | 95 | 81-105 |
| Mouse Serum (n=18) | 108 | 81-122 |
| Rat Cell Culture Supernates (n=4) | 114 | 103-125 |
| Rat EDTA Plasma (n=7) | 96 | 83-114 |
| Rat Serum (n=10) | 92 | 81-115 |
Linearity
To assess the linearity of the assay, four or more samples containing and/or spiked with high concentrations of mouse/rat Leptin in each matrix were diluted with Calibrator Diluent and then assayed.
Scientific Data Images for Mouse/Rat Leptin Quantikine ELISA Kit
Mouse/Rat Leptin ELISA Standard Curve
Mouse/Rat Leptin/OB ELISA Standard Curve
Preparation and Storage
Shipping
The product is shipped with polar packs. 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
Additional Leptin/OB Products
Product Documents for Mouse/Rat Leptin Quantikine ELISA Kit
Product Specific Notices for Mouse/Rat Leptin Quantikine ELISA Kit
For research use only
⚠ WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to www.P65Warnings.ca.gov.Citations for Mouse/Rat Leptin Quantikine ELISA Kit
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Sample Tested: SerumVerified Customer | Posted 04/03/2018
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Sample Tested: 7TD1 mouse hybridoma cell lineVerified Customer | Posted 06/05/2017
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Sample Tested: Mouse/Rat Leptin Elisa KitVerified Customer | Posted 06/01/2017
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Sample Tested: Purified mouse IL-1beta proteinVerified Customer | Posted 04/21/2017
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Protocols
View specific protocols for Mouse/Rat Leptin Quantikine ELISA Kit (MOB00):
Refer to the product for complete assay procedure.
Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
- Prepare all reagents, standard dilutions, and samples as directed in the product insert.
- Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.
- Add 50 µL of Assay Diluent to each well.
- Add 50 µL of Standard, Control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.
- Aspirate each well and wash, repeating the process 4 times for a total of 5 washes.
- Add 100 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.
- Aspirate and wash 5 times.
- Add 100 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.
- Add 100 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.
50 µL Assay Diluent
50 µL Standard, Control, or Sample
100 µL Conjugate
100 µL Substrate Solution
100 µL Stop Solution
Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.
- ELISA Sample Preparation & Collection Guide
- ELISA Troubleshooting Guide
- How to Run an R&D Systems DuoSet ELISA
- How to Run an R&D Systems Quantikine ELISA
- How to Run an R&D Systems Quantikine™ QuicKit™ ELISA
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- Sandwich ELISA (Colorimetric) – Biotin/Streptavidin Detection Protocol
- Sandwich ELISA (Colorimetric) – Direct Detection Protocol
- Troubleshooting Guide: ELISA
- View all Protocols, Troubleshooting, Illustrated assays and Webinars
FAQs for Mouse/Rat Leptin Quantikine ELISA Kit
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Q: Why is the acid activation procedure required for pregnant mouse serum samples from second trimester through post-partum day 1 in the Mouse/Rat Leptin Quantikine® ELISA assay?
A: Mouse Leptin R intereferes in the MOB00 assay at high concentration >2000 pg/mL, and studies have shown that there is a massive increase in the level of criculating Leptin Receptor (OB-R) during later stage of pregnancy. Acid activation helps to dissociate the interaction between Leptin and its high affinity recceptor OB-R thus minimizing possible interference in the Leptin assay. Note that activation is not required for samples obtained from non-pregnant mice. Ref. J. Biol. Chem (1997) 272(48):30546-51.
