Human IL-1 Beta/IL-1F2/IL1B ELISA Kit PicoKine™

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We haven't tested ek0392, ek0410 ek0525 , and ek0437 on colostrum/breast milk. We are uncertain if the kits are able to work on colostrum/breast milk. Please suggest the custome run pilot test if he wants to try. In theory, if the expression level of the targets protein in the colostrum/breast milk falls in the assay range of the kits, the kits are able to detect the protein in the sample.

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The suggested dilution for plasma samples is 1:1. EK0392 hasn't been tested for any tissues. Endogenous biotin in tissue homogenates might introduce false positive signal. We recommend the customer run a pilot test as described below to confirm the affect of endogeous biotin first. Add tissue homogenates into the wells and then add ABC abd TMB without adding any biotinylated detection antibody to see if any signal will be observed. If no signal is produced, the customer can work on the tissue sample by using the kit.

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Our lab technicans haven't woked on

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For the product EK0392 you will need a 100ul of antibody for each 96 well plate. So if you are going to use all the 96 well plate, you need 9600ul (9.6ml) volume of antibody. Usually there is a loss of few micro liters while pipetting, so we suggest preparing more than what it is required.

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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: unable to understand 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-1 beta 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: 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 depends upon the kit used so please refer to each datasheet for details.

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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 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. 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. 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 IL-1 beta ELISA kit to have sensitivity of 0.15pg/ml, that means the minimum amount of IL-1 beta that can be declared/interpreted as positive by the above standard is 0.15pg/ml.

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A: in 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 have an affect on immunoreactivity, so if lysate is not a validated sample type, care must be taken in sample preparation and validation.

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A: we do not recommend freezing 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-1 beta for a later time, you should let the blood clot in glass tubes and collect the serum to freeze for later use.

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This ELISA does not recognize specifically the mature form of IL1-beta.

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There is cross-reactivity with the IL1 beta precursor pro-IL1 beta.

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We haven't got any ELISA that is specific to the mature IL-1-beta.

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We haven't got any ELISA that is specific to the mature IL-1-beta.

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The Human IL-1 beta EK0392 Elisa Kit has not been tested on tissue homogenates. Endogenous biotin in tissue homogenates might introduce a false positive signal. We recommend to run a pilot test to confirm the effect of the endogenous biotin first. 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, proceed and work on the tissue sample by using the kit.