Human Leptin ELISA Kit PicoKine™

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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

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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.

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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.

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Please see the following recommended dilution ratios for Human Leptin PicoKine

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COA

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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.

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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.

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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.

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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.

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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.

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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.

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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.

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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.

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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.

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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.

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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.

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For serum and plasma (heparin, EDTA), the suggested dilution ratio from normal healthy human is 1:20.