Human Leptin ELISA Kit - Quantikine Best Seller
R&D Systems, part of Bio-Techne | Catalog # DLP00
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)
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
Reactivity
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
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.
Additional Leptin/OB Products
Product Documents for Human Leptin Quantikine ELISA Kit
Product Specific Notices for Human Leptin Quantikine ELISA Kit
For research use only
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⚠ 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.