Human TNF Alpha/Tumor necrosis factor ELISA Kit PicoKine™

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EK0525 is not suitable for working with whole cell lysates.

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The immunogen sequence of the antibodies in the kits is V77-L233 which is the soluble form of TNF alpha. The protein is synthesized in the intracellular space of cells and secretes into the extracellular space which is cell culture supernatant. And the kit might be able to detect patial intracellular secreted protein which hasn't released into extracellular space in cell lysate. The kit would work with Hep2G cell supernatant.

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Our product EK0525 Human TNF Alpha ELISA Kit is not specifically validated with urine (only serum, plasma, and cell culture supernatants). However, we do not have concentration levels to report. When we tested human serum with EK0525 Human TNF Alpha ELISA Kit we found the measured levels were below the sensitivity limit which is 20 pg/ml not 1pg/ml as specified in the protocol. Thus, if testing body fluids, we would recommend using EK0525 Human TNF Alpha ELISA Kit mainly for spiking and recovery studies which have expected elevated TNF-alpha levels than normal.

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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 TNF-alpha 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: 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 needed 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: 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 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: 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: Boster Bio does not assess sample stability. Variations in sample collection, processing, and storage may affect 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 in order to limit protein degradation.

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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 impact immunoreactivity, so if lysate is not a validated sample type, care must be taken in sample preparation and validation.

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A: please read this article on ELISA data analysis. bosterbio.com/ELISA-data-analysis-instructions. we also provide a convenient online tool that you can use to analyze ELISA data. bosterbio.com/biology-research-tools/ELISA-data-analysis-online

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The sensitivity is the lower limit of target protein that can be detected by the kit. It is determined by adding two standard deviations to the mean O.D. value of twenty (20) blank wells and calculating the corresponding concentration. At the sensitivity level, the kit can detect the presence of the protein but not accurately quantify it. Accurate quantitation begins at the lowest point on the standard curve. For the Human TNF Alpha ELISA Kit PicoKine™ (EK0525), the values lower than 15.6pg/ml might not be reliable. If the user would like to see the signal difference at small concentrations, ELISA kits with higher sensitivity provided by other vendors are suggested to be used.

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The Human TNF Alpha ELISA Kit PicoKine™ (EK0525) hasn't been tested with human gingival cervicular fluid.We suggest to run a pilot test for this. In theory, if the expression level of the target protein in the urine samples falls in the assay range of the kits, the kits are able to detect the protein in the sample.

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For human serum samples using the Human TNF Alpha ELISA Kit PicoKine™ (EK0525), dilution of 1:1 is recommended.

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Expiration is based on storage temperature. Store at 4°C for 6 months, at -20°C for 12 months. Avoid multiple freeze-thaw cycles.

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Yes, the Human TNF Alpha ELISA Kit PicoKine™ (EK0525) would work with Hep2G cell supernatant.

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The immunogen sequence of the antibodies in the Human TNF Alpha ELISA Kit PicoKine™ (EK0525) is V77-L233 which is the soluble form of TNF alpha. The protein is synthesized in the intracellular space of cells and secretes into the extracellular space which is cell culture supernatant. And the kit might be able to detect partial intracellular secreted protein which has not released into extracellular space in cell lysate.

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The assay range of the Human TNF Alpha ELISA Kit PicoKine™ (EK0525) human TNF alpha kit in cell culture supernates is 15.6pg/ml-1000pg/ml.

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The assay range of the Human TNF Alpha ELISA Kit PicoKine™ (EK0525) human TNF alpha kit in serum and plasma samples is 7.8pg/ml-500pg/ml.