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Caspase-3 Fluorometric Assay Kit

based on 72 citations in multiple journalsCaspase-3 Fluorometric Assay Kit724.7 5
Convenient, Highly-Sensitive Assay
Catalog #: K105
SKU-Size Size Price Qty
K105-25 25 assays
$145.00
K105-100 100 assays
$395.00
K105-200 200 assays
$525.00
K105-400 400 assays
$855.00
More Sizes Get Quote

Product Details

Detection Method Fluorecence (400 nm excitation filter and 505 nm emission filter)
Species Reactivity Mammalian
Applications Detect early/middle stages of apoptosis; differentiate apoptosis from necrosis.
Features & Benefits • Simple one-step procedure; takes 1-2 hours
• Fast and convenient
• Comparison of the fluorescence of AFC from an apoptotic sample with an uninduced control allows determination of the fold increase in caspase-3/CPP32 activity
Kit Components • Cell Lysis Buffer
• 2X Reaction Buffer
• DEVD-AFC (1 mM)
• DTT (1 M)
Storage Conditions -20°C
Shipping Conditions Gel Pack
USAGE For Research Use Only! Not For Use in Humans.

Details

Activation of ICE-family proteases/caspases initiates apoptosis in mammalian cells. The CPP32/Caspase-3 Fluorometric Protease Assay Kit provides a simple and convenient means for assaying the DEVD-dependent caspase activity. The assay is based on detection of cleavage of substrate DEVD-AFC (AFC: 7-amino-4-trifluoromethyl coumarin). DEVD-AFC emits blue light (λmax = 400 nm); upon cleavage of the substrate by CPP32 or related caspases, free AFC emits a yellow-green fluorescence (max = 505 nm), which can be quantified using a fluorometer or a fluorecence microtiter plate reader. Comparison of the fluorescence of AFC from an apoptotic sample with an uninduced control allows determination of the fold increase in caspase-3/CPP32 activity.


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Do the protease inhibitor that I added will interfere with the assay?
Protease inhibitors will interefere with the assay. Caspase 3 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-3. Inhibition of caspase 3 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 3 or cleaved caspase 3?
This kit will detect active Caspase-3 from samples. The procasepase-3 (inactive) is proteolytically processed into the active Caspase-3. 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.
Shahsavari, Zahra et al. (2016) RIP1K and RIP3K provoked by shikonin induce cell cycle arrest in the triple negative breast cancer cell line, MDA-MB-468: necroptosis as a desperate programmed suicide pathway, Tumour Biol. 2016 Apr;37(4):4479-91.
Luo, Yun et al. (2017) Elatoside C protects against ox-LDL-induced HUVECs injury by FoxO1-mediated autophagy induction, Biochim Biophys Acta. 2017 Jun;1863(6):1654-1665.
Hung et al., Homocysteine facilitates LOX-1 activation and endothelial death through the PKCβ and SIRT1/HSF1 mechanism: relevance to human hyperhomocysteinaemia. Clin. Sci., Jun 2015; 129: 477 - 487.
Liang et al., Integrinβ6-Targeted Immunoliposomes Mediate Tumor-Specific Drug Delivery and Enhance Therapeutic Efficacy in Colon Carcinoma. Clin. Cancer Res., Mar 2015; 21: 1183 - 1195.
Ramphul et al., Plasmodium falciparum evades mosquito immunity by disrupting JNK-mediated apoptosis of invaded midgut cells. PNAS, Feb 2015; 112: 1273 - 1280.
For more citations of this product click here