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Product Details
| Detection Method | Fluorescence microscopy, Flow cytometry and fluorescence plate reader (Ex. = 485 nm and Em. = 535 nm) |
|---|---|
| Species Reactivity | Mammalian |
| Applications | Sensitive detection of activated caspase-9 in living cells. |
| Features & Benefits | • Simple one-step procedure; takes only 1-2 hours • Fast and convenient • The FITC label allows for direct detection of the activated caspases in apoptotic cells |
| Kit Components | • FITC-LEHD-FMK • Wash Buffer • Z-VAD-FMK |
| Storage Conditions | -20°C |
| Shipping Conditions | Gel Pack |
| USAGE | For Research Use Only! Not For Use in Humans. |
Details
Activation of caspases plays a central role in apoptosis. The CaspGLOW™ Fluorescein Active Caspase-9 Staining Kit provides a convenient means for sensitive detection of activated caspase-9 in • Living cells. The assay utilizes the caspase-9 inhibitor LEHD-FMK conjugated to FITC (FITC-LEHD-FMK) as a fluorescent marker. FITC-LEHD-FMK is cell permeable, nontoxic, and irreversibly binds to activated caspase-9 in apoptotic cells.
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Can the kit be used with murine samples?
Yes, as long as the mice samples express active caspase-9, this kit can be used. Please refer to our product citations. The kit has been standardized for mammalian cells only.
Can we use this kit with paraffin or frozen tissue sections?
We do not recommend. Our tracers bind to active caspases, which are not likely to be active after paraffin fixation. Some reports have shown partial success with frozen sections.
Do different cell lines react differently or require different probe/tracer concentrations?
Yes. Some cell lines are more prone to high background levels of apoptosis, other cell lines require longer incubation periods and longer (and more thorough) wash steps due to decreased cell permeability.
Can this kit be used in fixed cells (1% PF)? Can you co-stain with an antibody at the same time as staining with this kit?
This kit for in-situ detection of active caspase-9. Fixing won’t keep the protein active. We have not tested co-staining with another antibody.
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.
Azad et al., Major Pathways of Polymyxin-Induced Apoptosis in Rat Kidney Proximal Tubular Cells. Antimicrob. Agents Chemother., Apr 2015; 59: 2136 - 2143.
Désio Aurélio Farias-de-Oliveira et al., Caspase-8 and caspase-9 mediate thymocyte apoptosis in Trypanosoma cruzi acutely infected mice. J. Leukoc. Biol., Feb 2013; 93: 227 - 234
Zhnag, X. et. al. Interferon-
Exacerbates Dry Eye–Induced Apoptosis in Conjunctiva through Dual Apoptotic Pathways. Invest. Ophthalmol. Vis. Sci., Aug 2011; 52: 6279 - 6285.
Exacerbates Dry Eye–Induced Apoptosis in Conjunctiva through Dual Apoptotic Pathways. Invest. Ophthalmol. Vis. Sci., Aug 2011; 52: 6279 - 6285.Jamadar-Shroff V et al (2009) Clin. Cancer Res.; 15: 1269 - 1276.
Ito, E. et al. Potential Use of Cetrimonium Bromide as an Apoptosis-Promoting Anticancer Agent for Head and Neck Cancer. Mol. Pharmacol. 2009; 76: 969-983.
For more citations of this product click here
