Human IL-7/Interleukin-7 ELISA Kit PicoKine™

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Our lab hasn't woked on placental tissue samples using EK0779 human IL-17 ELISA kit. We are uncertain if

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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-7 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: 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 required. 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 assessd 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 can be found in the product insert. Components in lysate and lysis buffer may affect 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 alter the stabilityof samples. It is recommend to assay sample on priority basis collection when possible, or aliquot into single use volumes and store samples frozen. limit repetitive freeze-thaw cycles with the stored samples to avoid protein degradation.

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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. in the example mentioned, 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 IL-7 ELISA kit to have sensitivity of 1pg/ml, that means the minimum amount of IL-7 that can be declared/interpreted as positive by the above standard is 1pg/ml.

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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: without having an understanding 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-7 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: 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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Our lab hasn't woked on placental tissue samples using Human IL-7 ELISA Kit PicoKine™ (EK0779). We are uncertain if cytokines can be detected accurately in tissue samples stored at room temperature in 70% ethanol. Endogenous biotin in tissue homogenates might introduce false positive signal. We recommend to 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, proceed and work on the tissue sample by using the kit.

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Our lab hasn't woked on placental tissue samples using Human IL-7 ELISA Kit PicoKine™ (EK0779). We recommend to 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, proceed and work on the tissue sample by using the kit. Please see the following suggested protocol: Preparation of lysate from tissues 1. Mince tissue and then rinse tissue in ice-cold PBS. 2. Place minced tissue in tissue homogenizer. 3. Add ice-cold PBS. For a 0.5g piece of tissue, add 5ml of ice-cold PBS(Volumes of PBS must be determined in relation to the amount of tissue present.) 5. Homogenize for several minutes at high speed. 6. Sonicate until no tissue chunks remain. 7. Centrifuge at ~10000 x g for 10 minutes. The centrifugation force and time can be varied depending on the sample type. 8. Aspirate supernatant. 9. Aliquot supernatant and store at -20°C.