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Product Details
| Cat # +Size | K160-100 |
|---|---|
| Size | 100 assays |
| Detection Method | Fluorescence (Ex/Em 400/505 nm) |
| Species Reactivity | Mammalian |
| Applications | Effective means for screening caspase inhibitors using fluorometric methods. |
| Features & Benefits | • Simple one-step procedure; takes only 1-1.5 hours • Fast and convenient • Inhibition of caspase-10 activity can be determined by comparison of the fluorescence intensity in samples with and without the testing inhibitors. |
| Kit Components | • 2X Reaction Buffer • Caspase Substrate AEVD-AFC (1 mM) • DTT (1 M) • Active Caspase-10 (Lyophilized) • Caspase Inhibitor, Z-VAD-FMK (2 mM) |
| Storage Conditions | -20°C |
| Shipping Conditions | Gel Pack |
| USAGE | For Research Use Only! Not For Use in Humans. |
Details
Caspases have been shown to play a crucial role in apoptosis induced by various deleterious and physiologic stimuli. Inhibition of caspases can delay apoptosis, implicating a potential role in drug screening efforts. The Caspase-10 Inhibitor Drug Screening Kit provides an effective means for screening caspase inhibitors using fluorometric methods. The assay utilizes synthetic peptide substrate AEVD-AFC (AFC, 7-amino-4-trifluoromethyl coumarin). Active caspase-10 cleaves the synthetic substrate to release free AFC which can then be quantified by fluorometry. Compounds to be screened can directly be added to the reaction and the level of inhibition of caspase-10 activity can be determined by comparison of the fluorescence intensity in samples with and without the testing inhibitors.
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Do the protease inhibitor that I added will interfere with the assay?
Protease inhibitors will interefere with the assay. Caspase 10 is a cysteine protease and can be inhibited by most protease inhibitor cocktails. The assay is based on the catalytic clevage of the substrate DEVD-pNa by Caspase 10. Inhibition of caspase 10 would thus interfere with the measurement.
Can you test plasma samples?
The kit is good for cell and tissue lysates only and not for plasma samples.
What is the sensitivity range?
The colorimetric substrate has a sensitivity range >1uM.
Will this kit detect full length caspase 10 or cleaved caspase 10?
This kit will detect active Caspase 10 from samples. The procaspase 10 (inactive) is proteolytically processed into the active Caspase 10. I hope this information helps.
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?
The expiry date is typically 1 year from the date of shipment
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.
