Mouse IL-17/IL-17A ELISA Kit PicoKine™

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We suggest when you reconstitute your standard, after it has been totally defrosted, spin the vial down, reconstitute with assay diluent and then spin down again before use. In making standard dilutions we recommend to keep everything on ice since diluted standard are not stable and need to be used at the same day. Mix your standard thoroughly by pipetting up and down but do not vortex the solution as this might damage the protein.

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The product EK0431, unfortunately the epitope mapping has not been performed, so we do not have any information at what exact region binds to the mouse IL-17 protein. But we can let you know that this antibody pair is binding between amino acids 26 and 158 of mature mouse IL-17 as the whole mature protein used as immunogen.

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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 IL-17 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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A: We have a web based ELISA curve fitting (4pl) and data analysis tool. Please give it a try: bosterbio.com/biology-research-tools/ELISA-data-analysis-online. You can also consult our article on ELISA data analysis: bosterbio.com/ELISA-data-analysis-instructions

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A: although the vials are bottled using aseptic techniques, heat-treated vials, and sterile stock solutions, they are not considered or guaranteed to be sterile. If sterile material is required for an experiment, the material can be filtered through a 0.2 micron filter designed for use with biological fluids.

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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: without knowing the physiological or pathological context of your samples we cannot recommend a dilution ratio without performing a pilot test with your samples. Here is how you can perform a pilot study on your own: perform a serial dilution of your samples on the IL-17 ELISA kit to make sure you have a linear ascending curve followed by a plateau, which signifies the samples saturating the detection limit of the kit. Then you can pick the dilution ratios from samples in the linear part of the curve as your experimental dilution ratio.
If you are interested in using our ELISA service, you can also send us your sample and we will take care of everything for you. You can check our service details here: bosterbio.com/services/assay-services/ELISA-testing-service
Since you mentioned you have limited samples, our cost effective multiplex ELISA service would fit perfectly for your needs, where we can generate dozens of data points using as little as 25ul sample volume. Information on this service is also in the above link.

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A: Chelating agents such as EDTA, Heparin and Citrate can irreversibly bind metal ions from the functional domain of IL-17 causing degradation of its protein structure. IL-17 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 IL-17 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: 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. a point of focus 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. normally, 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 changeing 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 IL-17 ELISA kit to have sensitivity of 1pg/ml, that means the minimum amount of IL-17 that can be declared/interpreted as positive by the above standard is 1pg/ml.

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The Mouse IL-17 ELISA Kit PicoKine EK0431 has not been validated for tissue homogenates therefore it is not guaranteed. For tissue homogenates, endogenous biotin should be considered. Endogenous biotin in tissue homogenates might introduce false positive signal. Please run the test as described below to confirm if there is Endogenous biotin in the tissue lysate samples using the ELISA kit. • Add tissue homogenates into the wells and then add ABC and TMB without adding any biotinylated detection antibody to see if any signal will be observed. • If no signal is produced, then you can work on the tissue sample by using the kit.