Human TNFsR I ELISA Kit PicoKine™

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The Human TNFsR I ELISA Kit PicoKine endotoxin level is <0.5EU/mg antibody and yes it is azide free. Costumers needs to optimize the conditions, as the culture condition is different. The assay was performed on human peripheral blood mononuclear cells, incubated with 10 ng/ml TNSF4 for 5-6 days in serum-containing media. Other cells were incubated with TNSF4 that has been first incubated with 5-15ng/ml at 37C for one hour before adding to the medium.

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EK0528 is used to detect soluble fragment. Our lab hasn't tested EK0528 with any cell lysate, so we do not recommend using EK0528 to test cell lysate samples.

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A: for those Picokine® ELISAs where cell or tissue lysate is a validated sample type, sample preparation instructions for lysate can be found in the product insert. Components in lysate and lysis buffer can effect immunoreactivity, so if lysate is not a validated sample type, care must be taken in sample preparation and validation.

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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 might differ 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. what you should focus on is whether your sample O.D. values are statistically significantly higher than your blank values. considering 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 proportion 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 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 TNF RI ELISA kit to have sensitivity of 1pg/ml, that means the minimum amount of TNF RI that can be declared/interpreted as positive by the above standard is 1pg/ml.

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A: we recommend you this article on ELISA data analysis. bosterbio.com/ELISA-data-analysis-instructions. Boster also provides a free tool that you can use to analyze ELISA data. bosterbio.com/biology-research-tools/ELISA-data-analysis-online

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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: 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: Chelating agents such as EDTA, Heparin and Citrate can bind metal ions from the functional domain of TNF RI causing degradation of its protein structure. TNF RI may be denatured as a result and may compromise the assay's measurements. The chilating sites could also be too close to the epitopes required for detection and block the antigen antibody reaction. We have tested the TNF RI ELISA, treating samples with a number of anticoagulants and decided that heparin, EDTA or citrate can be used for treatment of blood/plasma samples. Do not use other anticoagulents when collecting samples.