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.
Best Seller
Human Leptin Quantikine ELISA Kit
R&D Systems | Catalog # DLP00
Loading...
Key Product Details
Assay Length
3.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (100 µL), Serum (10 µL), EDTA Plasma (10 µL), Heparin Plasma (10 µL), Citrate Plasma (10 µL)
Sensitivity
7.8 pg/mL
Assay Range
15.6-1000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Citrate Plasma)
Loading...
Product Summary for Human Leptin Quantikine ELISA Kit
The Quantikine Human Leptin Immunoassay is a 3.5 hour solid phase ELISA designed to measure soluble human Leptin in cell culture supernates, serum, and plasma. It contains E. coli-expressed recombinant human Leptin and antibodies raised against the recombinant factor. This immunoassay has been shown to quantitate recombinant Leptin accurately. Results obtained measuring natural human Leptin showed dose-response curves that were parallel to the standard curves obtained using the recombinant Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for natural human 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
Human
Specificity
Natural and recombinant human Leptin
Cross-reactivity
< 0.5% cross-reactivity observed with available related molecules. < 50% cross-species reactivity observed with species tested.
Interference
Interference observed with 1 or more available related molecules.
Sample Values
Serum - Samples from apparently healthy volunteers were evaluated for the presence of
human Leptin in this assay. No medical histories were available for the donors used in this
study.
Five additional male serum samples fell below the lowest standard,15.6 pg/mL, when diluted 100-fold.
ND=Non-detectable
*Samples were dilluted 20-fold prior to assay.
| Sample Type | Range (pg/mL) | Mean (pg/mL) |
| Male Serum (n=16) | 2205-11,419 | 4760 |
| Female Serum (n=36) | 3877-77,273 | 20,676 |
Five additional male serum samples fell below the lowest standard,15.6 pg/mL, when diluted 100-fold.
Note: Values in EDTA and heparin plasma have been found to be comparable to paired serum
samples. Values in citrate plasma have been found to be slightly decreased compared to paired
serum, EDTA or heparin plasma samples.
Cell Culture Supernates:
Human peripheral blood mononuclear cells (5 x 106
cells/mL) were cultured in RPMI
supplemented with 5% fetal bovine serum, 50 μM beta -mercaptoethanol, 2 mM L-glutamine,
100 U/mL penicillin, and 100 μg/mL streptomycin sulfate. The cells were cultured unstimulated
or stimulated with 10 μg/mL PHA. Aliquots of the cell culture supernates were removed on
days 1 and 5 and assayed for levels of human Leptin.
| Condition | Day 1(pg/mL) | Day 5 (pg/mL) |
| Unstimulated | ND | ND |
| Stimulated | 152 | 75.9 |
BeWo human choriocarcinoma cells (1 x 106
cells/mL) were cultured in F-12 media
supplemented with 15% fetal bovine serum. The cells were cultured unstimulated or
stimulated with 2 μM forskolin and 20 μM forskolin. Aliquots of the cell culture supernates
were removed on days 1, 2, and 3 and assayed for levels of human Leptin.
| Condition | Day 1 (pg/mL) | Day 2 (pg/mL) | Day 3 (pg/mL) |
| Unstimulated* | 849 | 1549 | 1667 |
| 2 μM forskolin* | 1231 | 1699 | 2054 |
| 20 μM forskolin* | 1137 | 1725 | 2747 |
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, Citrate Plasma, EDTA Plasma, Heparin Plasma, Serum
| Intra-Assay Precision | Inter-Assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 40 | 40 | 40 |
| Mean (pg/mL) | 64.5 | 146 | 621 | 65.7 | 146 | 581 |
| Standard Deviation | 2.14 | 4.32 | 20.0 | 3.56 | 6.17 | 20.6 |
| CV% | 3.3 | 3.0 | 3.2 | 5.4 | 4.2 | 3.5 |
Recovery for Human Leptin Quantikine ELISA Kit
The recovery of Leptin spiked to three different levels throughout the range of the assay in various matrices was evaluated.
| Sample Type | Average % Recovery | Range % |
|---|---|---|
| Cell Culture Media (n=5) | 98 | 94-102 |
| Citrate Plasma (n=5) | 95 | 87-105 |
| EDTA Plasma (n=5) | 99 | 85-112 |
| Heparin Plasma (n=5) | 90 | 81-100 |
| Serum (n=5) | 95 | 89-109 |
Linearity
To assess the linearity of the assay, cell culture media samples spiked with high concentrations of Leptin and natural serum/plasma samples were diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Scientific Data Images for Human Leptin Quantikine ELISA Kit
Human Leptin/OB ELISA Standard Curve
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
Additional Leptin/OB Products
Product Documents for Human Leptin Quantikine ELISA Kit
Certificate of Analysis
To download a Certificate of Analysis, please enter a lot or batch number in the search box below.
Note: Certificate of Analysis not available for kit components.
Product Specific Notices for Human 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 Human Leptin Quantikine ELISA Kit
Powered by Bioz
Customer Reviews for Human Leptin Quantikine ELISA Kit (9)
4.9 out of 5
9 Customer Ratings
Have you used Human Leptin Quantikine ELISA Kit?
Submit a review and receive an Amazon gift card!
$25/€18/£15/$25CAN/¥2500 Yen for a review with an image
$10/€7/£6/$10CAN/¥1110 Yen for a review without an image
Submit a review
Customer Images
Showing
1
-
5 of
9 reviews
Showing All
Filter By:
-
Sample Tested: SerumVerified Customer | Posted 11/14/2022
-
Sample Tested: EDTA PlasmaVerified Customer | Posted 05/25/2022
-
Sample Tested: Serum and EDTA PlasmaVerified Customer | Posted 05/20/2022
-
Sample Tested: SerumVerified Customer | Posted 01/31/2022
-
Sample Tested: SerumVerified Customer | Posted 01/14/2022
-
Sample Tested: SerumVerified Customer | Posted 01/27/2021
-
Sample Tested: SerumVerified Customer | Posted 01/25/2021Per usual, R&D kit was easy to use and straight forward. Produced great results.
-
Sample Tested: EDTA PlasmaVerified Customer | Posted 01/22/2021Product preformed well- instructions were clear, included detailed background, and the reagent was easy to prepare.
-
Sample Tested: Human serumVerified Customer | Posted 05/15/2017
There are no reviews that match your criteria.
Protocols
View specific protocols for Human Leptin Quantikine ELISA Kit (DLP00):
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 100 µL of Assay Diluent to each well.
- Add 100 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 3 times for a total of 4 washes.
- Add 200 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 1 hour.
- Aspirate and wash 4 times.
- Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.
- Add 50 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.
100 µL Assay Diluent
100 µL Standard, Control, or Sample
200 µL Conjugate
200 µL Substrate Solution
50 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
- Quantikine HS ELISA Kit Assay Principle, Alkaline Phosphatase
- Quantikine HS ELISA Kit Principle, Streptavidin-HRP Polymer
- Sandwich ELISA (Colorimetric) – Biotin/Streptavidin Detection Protocol
- Sandwich ELISA (Colorimetric) – Direct Detection Protocol
- Troubleshooting Guide: ELISA
- View all Protocols, Troubleshooting, Illustrated assays and Webinars
