Rat KIM1 ELISA Kit PicoKine™

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EK0882 is not suitable for working on mouse samples?

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We did not test the cross reactivity of antibody from this kit to mouse protein. Therefore, we cannot guarantee the result if you use it to mouse.

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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 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 valid 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 check 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: 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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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. avoid repeat freeze-thaw cycles with the stored samples in order to prevent protein degradation.

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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 TIM-1 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 Picokine® ELISA Kits will generally run a 7-point standard curve, non-specific binding wells, and 39 samples in duplicate. this may vary slightly by kit so please refer to each datasheet for details.

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A: Chelating agents such as EDTA, Heparin and Citrate can irreversibly bind metal ions from the functional domain of TIM-1 causing degradation of its protein structure. TIM-1 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 TIM-1 ELISA, treating samples with different 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: 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 impact immunoreactivity, so if lysate is not a validated sample type, care must be taken in sample preparation and validation.

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For the Rat KIM1 ELISA Kit PicoKine™ (EK0882), the following points may help you: 1. Know that the ABC amount is redundant, so that having twice the amount of ABC in standard wells will only affect the readings marginally, skewing the results higher. 2. Having 50% of ABC in sample wells could be problematic if the samples have high amounts of KIM1 in them. Whether that is the case, you will have to decide based on the specific models you are using. If the level of KIM1 is low, it is expected to only marginally affect the readings, skewing the results lower. If KIM1 is high, and binds to more capture antibodies than there is ABC (at only 50% amount) to fully cover, then it would skew the results a lot. This is unlikely to happen but please check to see if your model has a very high level of KIM1. 3. With the same amount analytes generating higher readings in standard wells and lower readings in sample wells, your conclusion of the amount of KIM1 in each sample will be skewed lower than their actual value. That means, the deduced level of KIM1 is lower than the actual level of KIM1. Due to the redundancy in the amount of ABC, the deduced value should be closer to the actual value than the 50%. The data is usable as guidance for further experiment designs but is likely not acceptable for publication. The above are our technical opinions. Please take in considerations of other circumstances that we are not aware of and make a comprehensive decision.

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The Rat KIM1 ELISA Kit PicoKine™ (EK0882) is not suitable for working on mouse samples. Our lab hasn't worked on tissue samples using the kit. 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, you can proceed and work on the tissue sample by using the kit.