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Product Info Summary
| SKU: | EK0437 |
|---|---|
| Size: | 96 wells/kit, with removable strips. |
| Reactive Species: | Human |
| Application: | ELISA |
| Sample Types: | cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA). |
Product info
Product Name
Human Leptin ELISA Kit PicoKine®
SKU/Catalog Number
EK0437
Size
96 wells/kit, with removable strips.
*Question: How many samples can I assay/run in this kit?
Description
Human Leptin ELISA Kit PicoKine™ (96 Tests). Quantitate Human LEP in cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA). Sensitivity: 10pg/ml.
Storage & Handling
Store at 4°C for 6 months, at -20°C for 12 months. Avoid multiple freeze-thaw cycles (Ships with gel ice, can store for up to 3 days in room temperature. Freeze upon receiving.)
Cite This Product
Human Leptin ELISA Kit PicoKine® (Boster Biological Technology, Pleasanton CA, USA, Catalog # EK0437)
Clonality of Antibodies
See Datasheet for details
Immunogen
Expression system for standard: E.coli; Immunogen sequence: V22-C167
Sensitivity
<10 pg/ml
Assay Range
62.5 pg/ml - 4,000 pg/ml
Standard Dilution Instructions
Add 100ul of sample diluent in well #2-#8. Add 200ul standard stock solution to well #1, and serial dilute for well #2-#7 to make a standard curve row. Leave well #8 as blank See datasheet of EK0437 for more details
Cross-reactivity
There is no detectable cross-reactivity with other relevant proteins.
Reactive Species
EK0437 is reactive to LEP in Human samples
Validated Sample Types
cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA).
Application Guarantee
EK0437 is guaranteed for ELISA in Human by Boster Guarantee
See how Boster Bio validate our ELISA kits: ELISA Validation Information
Background of Leptin/OB
Leptin (or obese, OB) is a circulating hormone that is expressed abundantly and specifically in the adipose tissue. Human ob protein is a 166-amino acid polypeptide with a putative signal sequence and is 84 and 83% homologous to the mouse and rat ob proteins, respectively. Northern blot analysis reveals that OB RNA is present at a high level in adipose tissue but at much lower levels in placenta and heart. Indeed, Leptin is first described as an adipocyte-derived signalling factor, which, after interaction with its receptors, induces a complex response including control of bodyweight and energy expenditure. Leptin seems in addition to its role in metabolic control to have important roles in reproduction and neuroendocrine signalling. Leptin is a powerful inhibitor of bone formation in vivo. Comparative mapping of mouse and human DNA indicates that the ob gene is located within a region of mouse chromosome 6 that is homologous to a portion of human chromosome 7q. The standard product used in this kit is recombinant human Leptin.
Kit Components
| Catalog Number | Description | Quantity |
|---|---|---|
| EK0437-CAP | Anti-Human LEP Pre-coated 96-well strip microplate | 1 |
| EK0437-ST | Human LEP Standard | 2 vials, 10 ng/tube |
| EK0437-DA | Human LEP Biotinylated antibody (100x) | 100ul |
| AR1103 | Avidin-Biotin-Peroxidase Complex (100x) | 100ul |
| AR1106-1 | Sample Diluent | 30ml |
| AR1106-2 | Antibody Diluent | 12ml |
| AR1106-3 | Avidin-Biotin-Peroxidase Diluent | 12ml |
| AR1104 | Color Developing Reagent (TMB) | 10ml |
| AR1105 | Stop Solution | 10ml |
| AR1106-5 | Wash Buffer (25x) | 20ml |
| PLA-SEA | Adhesive plate sealers | 4 |
*The kit components are not available for individual purchase.
Materials Required But Not Included In Kit
- Microplate Reader capable of reading absorbance at 450nm.
- Incubator.
- Automated plate washer (optional).
- Pipettes and pipette tips capable of precisely dispensing 0.5 µl through 1 ml volumes of aqueous solutions.
- Multichannel pipettes are recommended for large amount of samples.
- Deionized or distilled water.
- 500ml graduated cylinders.
- Test tubes for dilution.
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Data Examples, Quality Control Data & Sample Dilution
Validation Standard Curve O.D. At 450nm
| Concentration (pg/ml) | 0 | 62.5 | 125 | 250 | 500 | 1000 | 2000 | 4000 |
| O.D. | 0.002 | 0.015 | 0.036 | 0.080 | 0.191 | 0.505 | 0.923 | 1.801 |
Data Example Images
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Human Leptin PicoKine ELISA Kit standard curve
Recommended Sample Dilution Ratios
According to our internal validation assays using this ELISA kit, to detect Leptin/OB, Dilution ratio of 1:10, concentration in serum and plasma is 300-16000 pg/ml..
Some articles we found to cite concentrations of Leptin/OB in samples: 24900992 (Pubmed IDs).
Intra Assay Consistency & Inter Assay Consistency
We measured random samples of Human Leptin ELISA Kit PicoKine® within the same batch/lot to ensure the consistency of the kits' performances. ELISA intra assay consistency is measured using wells from the same plate/assay kit. ELISA inter assay consistency is measured using wells from different plates from the same batch production/lot.
| Intra-Assay Precision | Inter-Assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 16 | 16 | 16 | 24 | 24 | 24 |
| Mean (pg/ml) | 81 | 485 | 1057 | 75 | 444 | 996 |
| Standard deviation | 6.15 | 32.49 | 54.96 | 8 | 35.52 | 69.72 |
| CV (%) | 7.6% | 6.7% | 5.2% | 8.4% | 8% | 7% |
Reproducibility
We ensure reproducibility by testing three samples with differing concentrations of Leptin/OB in ELISA kits from four different production batches/lots.
| Lots | Lot 1 (pg/ml) | Lot 2 (pg/ml) | Lot 3 (pg/ml) | Lot 4 (pg/ml) | Mean (pg/ml) | Standard Deviation | CV (%) |
|---|---|---|---|---|---|---|---|
| Sample 1 | 81 | 83 | 84 | 87 | 83 | 2.16 | 2.6% |
| Sample 2 | 485 | 539 | 463 | 476 | 490 | 28.93 | 5.9% |
| Sample 3 | 1057 | 1101 | 1114 | 1116 | 1097 | 23.8 | 2.1% |
Protein Target Info & Infographic
Gene/Protein Information For LEP (Source: Uniprot.Org, NCBI)
Gene Name
LEP
Full Name
Leptin
Weight
18.641kDa
Superfamily
leptin family
Alternative Names
FLJ94114; LEP; leptin (murine obesity homolog); leptin (obesity homolog, mouse); Leptin; OB; Obese protein; obese, mouse, homolog of; Obesity factor; OBOBS LEP LEPD, OB, OBS leptin leptin|leptin (murine obesity homolog)|leptin (obesity homolog, mouse)|obese protein|obese, mouse, homolog of|obesity factor
*if product is indicated to react with multiple species, protein info is based on the gene entry specified above in "species".For more info on LEP, check out the LEP Infographic
We have 30,000+ of these available, one for each gene! check them out.
In this infographic you will see the following information for LEP: database IDs, super-family, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact us [email protected].
Specific Publications For Human Leptin ELISA Kit PicoKine® (EK0437)
Hello CJ!
EK0437 has been cited in 44 publications:
*The publications in this section are manually curated by our staff scientists. They may differ from Bioz's machine gathered results. Both are accurate. If you find a publication citing this product but is missing from this list, please let us know we will issue you a thank-you coupon.
The Relationship between Serum Zonulin Level and Clinical and Laboratory Parameters of Childhood Obesity
Metabolic Differences between Subcutaneous and Visceral Adipocytes Differentiated with an Excess of Saturated and Monounsaturated Fatty Acids
Artificial Intelligence-Assisted Identification of Genetic Factors Predisposing High-Risk Individuals to Asymptomatic Heart Failure
Leptin Overexpression in Bone Marrow Stromal Cells Promotes Periodontal Regeneration in a Rat Model of Osteoporosis
Elective percutaneous coronary intervention leads to significant changes in serum resistin, leptin, and adiponectin levels regardless of periprocedural myocardial injury: an observational study
Oxidative stress and adipokine levels were significantly correlated in diabetic patients with hyperglycemic crises
Perilipin polymorphisms are risk factors for the development of obesity in adolescents? A case-control study
The Effect of Isotretinoin on Retinol-Binding Protein 4, Leptin, Adiponectin and Insulin Resistance in Acne Vulgaris Patients
Cardiometabolic risks vary by weight status in pediatric kidney and liver transplant recipients: A cross‐sectional, single‐center study in the USA
Leptin concentration and oxidative stress in diabetic ketoacidosis
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Customer Reviews
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Customer Q&As
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Find answers in Q&As, reviews.
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Submit your question
17 Customer Q&As for Human Leptin ELISA Kit PicoKine®
Question
Do you have any information available documenting the effects of aprotinin on the ELISA results using EK0437?
Verified customer
Asked: 2020-12-01
Answer
Our lab hasn't validated effects of aprotinin on the ELISA results using the Human Leptin ELISA Kit PicoKine™ (EK0437). Please run a pilot test to see if it has any effect.
Boster Scientific Support
Answered: 2020-12-01
Question
We have acquired the products leptin (EK0437), adiponectin (EK0595) and endothelin (EK0945) from BOSTER and our sample type is humans' plasma (EDTA). We have read the respective datasheets. Even so, we need helping, since we have not conditions to test sample dilution proportion. What dilution ratio do you suggest for using with these kits? 1:10, 1:100, 1:1000 or another one? Please, give us numbers.
Verified Customer
Verified customer
Asked: 2020-02-28
Answer
Please see the following recommended dilution ratios for plasma (EDTA) from normal healthy human. leptin (EK0437), 1:20 adiponectin (EK0595) around 1:2000 endothelin (EK0945) 1:1
Boster Scientific Support
Answered: 2020-02-28
Question
Dear Technical Support: I am interested with Human Leptin ELISA Kit PicoKine for human population. My query is; how does these Human Leptin ELISA Kit PicoKine work well for urine sample in human adolescent? Is it required to concentrate the sample in order to measure the hormone?
Verified Customer
Verified customer
Asked: 2020-01-27
Answer
The product Human Leptin ELISA Kit PicoKine has not been tested particularly in human adolescents, however the typical human urine does not need to be concentrated and can be diluted using these Human Leptin ELISA Kit PicoKine if necessary.
Boster Scientific Support
Answered: 2020-01-27
Question
Q: if the enzyme conjugated Leptin antibodies are mixed with the substrate, will that convert the substrate into the enzymatic reaction product? Or the enzyme function is only activated when the antibody is attached with the Leptin antigen?
R. Allen
Verified customer
Asked: 2019-12-05
Answer
A: The enzyme is always active. Avoid contaminating the substrate with enzyme prior to the incubation otherwise it compromises the assay with false positive signal.
Boster Scientific Support
Answered: 2019-12-05
Question
Q: how many samples can be assayed in a Picokine® ELISA Kit?
Verified Customer
Verified customer
Asked: 2019-10-11
Answer
A: The Picokine® ELISA Kits will generally run a 7-point standard curve, non-specific binding wells, and 39 samples in duplicate. this may vary slightly by kit so please refer to each datasheet for details.
Boster Scientific Support
Answered: 2019-10-11
Question
What is the suggested dilution ratio for cell culture supernates using the Human Leptin ELISA Kit PicoKine™ (EK0437)?
Verified customer
Asked: 2019-09-19
Answer
For cell culture supernates, concentrations of the samples can vary greatly. We suggest to run pilot tests for different group samples to figure out the optimal dilution ratio. Our lab tested unstimulated human peripheral blood mononuclear cells with a dilution of 1:1 and didn't detect any measurable value.
Boster Scientific Support
Answered: 2019-09-20
Question
What is the suggested dilution ratio for cell lysates using the Human Leptin ELISA Kit PicoKine™ (EK0437)?
Verified customer
Asked: 2019-09-19
Answer
For cell lysates, concentrations of the samples can vary greatly. We suggest to run pilot tests for different group samples to figure out the optimal dilution ratio. Our lab tested unstimulated human peripheral blood mononuclear cells with a dilution of 1:1 and didn't detect any measurable value.
Boster Scientific Support
Answered: 2019-09-20
Question
What is the suggested dilution ratio for serum and plasma (heparin, EDTA) using the Human Leptin ELISA Kit PicoKine™ (EK0437)?
Verified customer
Asked: 2019-09-19
Answer
For serum and plasma (heparin, EDTA), the suggested dilution ratio from normal healthy human is 1:20.
Boster Scientific Support
Answered: 2019-09-20
Question
I want to purchase your Human Leptin ELISA Kit PicoKine, but I have some questions; does this kit would be useful in rabbit serum?
Verified Customer
Verified customer
Asked: 2019-08-15
Answer
The product Human Leptin ELISA Kit PicoKine have not been tested for any cross reactivity with rabbit samples, thus we do not know if the kit will work with rabbit serum samples. In this kit both capture and detection antibody are monoclonal so unless the homology between human and rabbit is very high, there would be a possibility that it will cross react, but we cannot guarantee.
Boster Scientific Support
Answered: 2019-08-15
Question
Q: What is the optimal O.D. value for Leptin ELISA kit? I used your Leptin ELISA on serum samples. For my positive control, I received an O.D. value of 0.826, while my negative control received a value of 0.136. I obtained both of these controls from the ELISA kit, where your kit's typical data shows O.D. values much higher than my positive control and your background is lower. My samples O.D. values are around 0.225 and the highest is only 0.357. can I consider these samples contain Leptin even though the O.D. values are not very high?
Verified Customer
Verified customer
Asked: 2019-06-20
Answer
A: The absolute O.D. values may change according to incubation time. The more you incubate the higher the O.D. values are going to be. an important assessment should be is whether your sample O.D. values are statistically significantly higher than your blank values. regarding your assay, you could extend your development time in the substrate incubation step to obtain higher O.D. values, as long as your negative controls' O.D. values are not increasing faster in relation to your positive controls. typically, a sample with O.D. value 2 standard deviations higher than your negative controls can be considered positive. We calculate the sensitivity of this ELISA kit by converting cutoff O.D. value, calculated as the average of 20 negative controls plus 2 standard deviations of the 20 negative controls, into a concentration. in other words, when we claim this Leptin ELISA kit to have sensitivity of 10pg/ml, that means the minimum amount of Leptin that can be declared/interpreted as positive by the above standard is 10pg/ml.
Boster Scientific Support
Answered: 2019-06-20
Question
Please help with this. On page 6 of the attached insert...it states the dilution of sample must be made with the sample diluent. But it does not state the target dilutions to be prepared by the user. I looked through this on the attached insert but also cannot understand. Please clarify for me asap Sample Dilution The target protein concentration should be estimated and appropriate sample dilutions should be selected such that the final protein concentration lies near the middle of the linear dynamic range of the assay. It is recommended to prepare 150 ?l of sample for each replicate to be assayed. The samples should be diluted with sample diluent and mixed gently.
Verified Customer
Verified customer
Asked: 2019-06-07
Answer
Please see the following recommended dilution ratios for Human Leptin PicoKine
Boster Scientific Support
Answered: 2019-06-07
Question
Q: Can Leptin ELISA Kits be used with tissue homogenates (or other non-validated sample types)?
S. Garcia
Verified customer
Asked: 2019-05-29
Answer
A: Unfortunately, Boster Bio has not routinely validated tissue homogenates as a sample type for ELISA kits. This does not mean that ELISA kits are not suitable for other sample types than we have tested: it means further investigation is a must. One will need to perform a spike and recovery study to determine if an unvalidated sample type will work with a particular kit. To perform a spike and recovery experiment, one should divide a sample into two aliquots. In one of the aliquots, the user should spike in a known amount of the kit standard. a dilution series is performed comparing the spiked versus the unspiked sample. Generally, samples with expected recovery and linearity between 80-120% are considered acceptable. This method can be used to validate any sample type that has not been evaluated by Boster Bio. for a more detailed spike and recovery protocol, please contact technical support.
Note: acceptable ranges should be determined individually by each laboratory. Additionally, technical support can help determine if a buffer component is not compatible with a given ELISA kit. please see the Citations tab on the product webpage for peer-reviewed papers utilizing a wide range of sample types. We also have an innovator's reward program where if the user validates our ELISA kits in applications or samples previously not validated by Boster Bio or other users, and share such information with us by submit a review, we will reward the user's efforts with a free antibody or ELISA kit from our catalog. Biocompare.com will also give $20 Amazon giftcard as an additional reward, if the review is submitted there as well.
Boster Scientific Support
Answered: 2019-05-29
Question
Q: what is the protocol regarding preparation of cell lysates prepared for use in Picokine® ELISA kits?
Verified Customer
Verified customer
Asked: 2019-01-14
Answer
A: for those Picokine® ELISAs where cell or tissue lysate is a validated sample type, sample preparation instructions for lysate are present in the product insert. Components in lysate and lysis buffer can change immunoreactivity, so if lysate is not a validated sample type, care must be taken in sample preparation and validation.
Boster Scientific Support
Answered: 2019-01-14
Question
Q: how long can samples (cell cultures, serum, and plasma) be stored and still be stable for analysis Leptin using the EK0437 Human Leptin Picokine® ELISA Kit?
D. Varadkar
Verified customer
Asked: 2018-09-19
Answer
A: Boster Bio does not evaluate sample stability. Variations in sample collection, processing, and storage may change the stabilityof samples. It is recommend to assay sample right after collection when possible, or aliquot into single use volumes and store samples frozen. avoid repetitive freeze-thaw cycles with the stored samples to prevent protein degradation.
Boster Scientific Support
Answered: 2018-09-19
Question
Q: can I use citrate plasma as samples in Human Leptin Picokine® ELISA Kit (Catalog # EK0437)?
Verified Customer
Verified customer
Asked: 2018-04-16
Answer
A: Chelating agents such as EDTA, Heparin and Citrate can attach metal ions from the functional domain of Leptin causing degradation of its protein structure. Leptin may be denatured as a result and may compromise the assay's measurements. The chilating sites could also be too close to the epitopes a must for detection and block the antigen antibody reaction. We have tested the Leptin ELISA, treating samples with various anticoagulants and decided that heparin or EDTA can be used for treatment of blood/plasma samples. Do not use other anticoagulents when collecting samples.
Boster Scientific Support
Answered: 2018-04-16
Question
Q: which procedure should I follow in order to thaw whole blood sample for Leptin ELISA after freezing?
Verified Customer
Verified customer
Asked: 2016-06-17
Answer
A: do not freeze and thaw whole blood. erythrocytes are fragile and, if frozen and thawed, will undergo hemolysis rendering the samples useless. To keep your blood samples to test Leptin for a later time, you should let the blood clot in glass tubes and separate the serum to freeze for later analysis.
Boster Scientific Support
Answered: 2016-06-17
Question
EK0437 Human Leptin PicoKine ELISA Kit I have a customer who is interested in the kit. They want to test it with human serum samples. Please provide your QC data for the current lot of kit along with reference values for serum samples.
Verified Customer
Verified customer
Asked: 2015-05-19
Answer
COA
Boster Scientific Support
Answered: 2015-05-19
