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CaspGLOW™ Fluorescein Active Caspase-8 Staining Kit

based on 20 citations in multiple journalsCaspGLOW™ Fluorescein Active Caspase-8 Staining Kit205 5
Detects activated Caspase-8 by FACS, FL or plate reader
Catalog #: K188
SKU-Size Size Price Qty
K188-25 25 assays
$175.00
K188-100 100 assays
$460.00
More Sizes Get Quote

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-8 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-IETD-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-8 Staining Kit provides a convenient means for sensitive detection of activated caspase-8 in • Living cells. The assay utilizes the caspase-8 inhibitor, IETD-FMK, conjugated to FITC (FITC-IETD-FMK) as a fluorescent in situ marker. FITC-IETD-FMK is cell permeable, nontoxic, and irreversibly binds to activated caspase-8 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-8, 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-8. 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.
I have tried to label apoptotic cells by caspase inhibitors and I do not know if K188 is appropriate for my samples. I would appreciate your advice.
K188 already uses the caspase-8 inhibitor, IETD-FMK, conjugated to FITC (FITC-IETD-FMK) as a fluorescent in situ marker. FITC-IETD-FMK binds to activated caspase-8 irreversibly and will be detected in apoptotic cells. If your caspase-8 inhibitor is already fluorescently labeled, then you don’t need K188. Your labeled inhibitor should be able to bind activated caspase-8 in apoptotic cells and then can be detected by Flow Cytometry or Fluorescence Microscopy.
Azad et al., Major Pathways of Polymyxin-Induced Apoptosis in Rat Kidney Proximal Tubular Cells. Antimicrob. Agents Chemother., Apr 2015; 59: 2136 - 2143.
Boulianne et al., AID and Caspase 8 Shape the Germinal Center Response through Apoptosis. J. Immunol., Dec 2013; 191: 5840 - 5847.
Adrienne Anginot et al., Galectin 1 Modulates Plasma Cell Homeostasis and Regulates the Humoral Immune Response. J. Immunol., Jun 2013; 190: 5526 - 5533.
Yang, L. et. al. Contributions of TRAIL-mediated megakaryocyte apoptosis to impaired megakaryocyte and platelet production in immune thrombocytopenia. Blood, Nov 2010; 116: 4307 - 4316.
Malignancies.  Clin. Cancer Res., 2010; 16: 3648-3658.
For more citations of this product click here