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TACE Inhibitor Screening Assay Kit ( Fluorometric)

Sensitive Assay, HTS
Catalog #: K366

Product Details

Cat # +Size K366-100
Size 100 assays
Detection Method Fluorescence (Ex/Em 318/449 nm)
Species Reactivity Mammalian
Applications Provides a rapid, simple, sensitive and reliable test suitable for high-throughput screening of TACE inhibitors and can be modified to check the relative TACE activity. Inhibitor control GM6001 is included to compare the efficiency of test inhibitors.
Features & Benefits • Simple procedure
• Fast and convenient
• The assay is sensitive, simple, and high-throughput adaptable.
Kit Components • TACE Assay Buffer
• TACE Substrate
• TACE Enzyme (20 μg)
• Inhibitor Control (0.1 mM GM6001)
Storage Conditions -20°C
Shipping Conditions Gel Pack
USAGE For Research Use Only! Not For Use in Humans.


The TACE (tumor necrosis factor-α-converting enzyme), also called ADAM metallopeptidase domain 17 (ADAM17), is a 70-kDa enzyme that belongs to the ADAM protein family of disintegrins and metalloproteases. TACE is believed to be involved in the processing of tumor necrosis factor alpha (TNF-α) at the surface of the cell, and from within the intracellular membranes of the trans-Golgi network. This process, which is also known as 'shedding', involves the cleavage and release of a soluble ectodomain from membrane-bound pro-proteins (such as pro-TNF-α), and is of known physiological importance. In BioVision’s TACE inhibitor screening Kit, TACE hydrolyzes the specific FRET substrate to release the quenched fluorescent group, which can be detected fluorometrically at Ex/Em = 318/449 nm. In the presence of the potent TACE inhibitor, the hydrolyzation of substrate will be impeded. The kit provides a rapid, simple, sensitive and reliable test suitable for high-throughput screening of TACE inhibitors and can be modified to check the relative TACE activity. Inhibitor Control GM6001 is included to compare the efficacy of test inhibitors.

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How can we measure TACE activity rather than the inhibitor screening using this kit?
There is no Standard in K366. Therefore, the absolute activity of the enzyme can’t be measured. However, the relative activity of TACE can be measured between Control vs treated samples and the fold difference can be calculated.
I have recently acquired the TACE Inhibitor screening kit. I want to use this kit to verify inhibitor activity in conditions as close to my standard assay system as possible. Thus, I would need to use incubate enzyme and substrate in cell-culture medium (if possible even with DMEM+FBS or cell-culture supernatant). Please tell me if this is possible, or if the kit only works with the supplied assay buffer.
This protocol has been designed for working with the assay buffer. To make it work with the cell culture media, some optimization may be needed by the end user. We have not tested this ourselves, so I cannot comment on whether it will be successful or not. The one suggestion I can give the client is to dilute whatever system he plans on using with our assay buffer so that all the ingredients required for the reaction to occur are present in the well.
How much enzyme is in the vial?
We give you 20 µg of it in every kit.
Do you happen to know what species the enzyme is?
This is a human recombinant enzyme.
Can this kit be used with samples like bacteria, plants, drosophila, yeast etc?
We have optimized the kit with mammalian samples. However, theoretically these kits should work with samples from multiple species/sources. Since the optimal conditions depend on the sample type, the protocol has to be be adapted to fit the samples for efficient results. Please refer to this kit's citations to see what kind of samples have been used with this kit other than mammalian samples.
Can we use frozen samples with this assay?
Fresh samples are always preferred over frozen samples. However, frozen samples can also be used, provided, they were frozen right after isolation, were not freeze thawed multiple time (for which we recommend aliquoting the samples before freezing) and have been frozen for relatively short periods.
Can we use a different wavelength than recommended for the final analysis?
It is always recommended to use the exact recommended wavelength for the most efficient results. However, most plate readers have flexibility in their band width of detection in increments of +/- 10 nm. Depending on this flexibility range, you can deviate from the recommended wavelengths within limits.
What is the exact volume of sample required for this assay?
There is no specific volume we can recommend for the amount any sample to be used since it is completely sample concentration and quality based. You have to do a pilot expt with multiple sample volumes to determine the optimal volume which gives a reading within the linear range of the standard curve. Please refer to the citations for this product to see what other clients have used with similar sample types.
Do you have trial sizes of this kit?
Unfortunately, we do not have trial sizes of this kit available. However, if you are based in the US or Canada, we will give you a 10% off list price introductory discount on its purchase price. If you are based out of this area please contact yopur regional BioVision distributor.
What is the shelf life of this kit?
This kit is good for 12 months from the date of shipment in the unopened form when stored at the appropriate temperature and appropriate conditions. After opening and reconstitution, some of the components in this kit are good for 2 months at -20°C. Please refer to the datasheet for storage information and shelf life of each of the components.
Why are my standard curve values lower than those shown on the datasheet?
There are multiple factors which influence the signals like the incubation times, room temperature, handling etc. In general, to increase the value of the standards, you can increase the incubation time. As long as the standard curve is linear, it should be fine to use, since all of your samples will also be measured under the same conditions on this curve.
How do I normalize my samples against protein concentration?
You can use a protein quantitation assay on the supernatants you get from cell/tissue lysates or with any other liquid sample in the assay buffer.
Can we purchase individual components of this kit?
Yes, you can purchase any of the kit's components without the whole kit. Please refer to the component Cat #s mentioned on the datasheet for ordering.
Can we use an alternate buffer for sample preparation (cell lysis, sample dilutions etc)?
Our assay buffers are optimized for the reactions they are designed for. They not only contain some detergents for efficient lysis of your cells/tissue, but also contain some proprietary components required for the further reactions. Therefore, we highly recommend using the buffers provided in the kit for the best results.
Should I make a standard curve for every expt I do, or is one curve/kit enough?
Yes, I would strongly recommend you to do the standards every time you do the expt. There is always a chance that something was done differently that day and we do not want any conditions to differ between standards and samples.
Does any of the kit components contain recombinant proteins developed using the baculoviral expression system?
The TACE enzyme is a human recombinant enzyme produced in insect S9 cells using a baculoviral expression system.
Do you have the information regarding what sample types (e.g. cell cultures, supernatant, tissue types …etc.) can be assayed with this kit? And what species of mammals (e.g. mouse, rats, human… etc.) did you checked for reactivity?
There is a fundamental misunderstanding here. The TACE enzyme provided in the kit can be tested with any sample containing putative inhibitors irrespective of species. There is no antibody involved and no species reactivity applies to this kit. We mention mammalian because that is the most common sample origin (includes cell culture, tissue/fluids etc.) for testing inhibitors.