Caspase 10 Fluorometric Assay Kit, AEVD
Description:
Caspase 10 Fluorometric Assay Kit, AEVD
Trade Name:
Chemicon (Millipore)
Qty/Pk:
25 assays
Product Overview:
Activation of ICE-family proteases/caspases initiates apoptosis in mammalian cells. Chemicon's Caspase-10 Fluorometric Protease Assay Kit provides a simple and convenient means for assaying the activity of caspases that recognize the sequence AEVD. The assay is based on detection of cleavage of substrate AEVD-AFC (AFC: 7-amino-4-trifluoromethyl coumarin). AEVD-AFC emits blue light (lmax=400 nm); upon cleavage of the substrate by caspase-10 or related caspases, free AFC emits a yellow-green fluorescence (lmax=505 nm), which can be quantified using a fluorometer or a fluorescence microtiter plate reader. Comparison of the fluorescence of AFC from an apoptotic sample with an uninduced control allows determination of the fold increase in AEVD-dependent caspase activity.
Key Applications:
Activity Assay
Species Reactivity:
Mammals
Usage Statement:
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
Entrez Gene Summary:
This gene encodes a protein which is a member of the cysteine-aspartic acid protease (caspase) family. Sequential activation of caspases plays a central role in the execution-phase of cell apoptosis. Caspases exist as inactive proenzymes which undergo proteolytic processing at conserved aspartic residues to produce two subunits, large and small, that dimerize to form the active enzyme. This protein cleaves and activates caspases 3 and 7, and the protein itself is processed by caspase 8. Mutations in this gene are associated with apoptosis defects seen in type II autoimmune lymphoproliferative syndrome. Three alternatively spliced transcript variants encoding different isoforms have been described for this gene.
UniProt Summary:
FUNCTION: SwissProt: Q92851 # Isoform C is proteolytically inactive.
SIZE: 521 amino acids; 58951 Da
SUBUNIT: Heterotetramer that consists of two anti-parallel arranged heterodimers, each one formed by a 23/17 kDa (p23/17) (depending on the splicing events) and a 12 kDa (p12) subunit (By similarity). Self-associates. Interacts with FADD and CASP8. Found in a Fas signaling complex consisting of FAS, FADD, CASP8 and CASP10.
TISSUE SPECIFICITY: Detectable in most tissues. Lowest expression is seen in brain, kidney, prostate, testis and colon.
PTM: Cleavage by granzyme B and autocatalytic activity generate the two active subunits. & Phosphorylated upon DNA damage, probably by ATM or ATR.
DISEASE: SwissProt: Q92851 # Defects in CASP10 are the cause of autoimmune lymphoproliferative syndrome type 2A (ALPS2A) [MIM:603909]. ALPS2 is characterized by abnormal lymphocyte and dendritic cell homeostasis and immune regulatory defects. & Defects in CASP10 are a cause of familial non-Hodgkin lymphoma (NHL) [MIM:605027]. NHL is a cancer that starts in cells of the lymph system, which is part of the body's immune system. NHLs can occur at any age and are often marked by enlarged lymph nodes, fever and weight loss. & Defects in CASP10 are a cause of gastric cancers [MIM:137215].
SIMILARITY: SwissProt: Q92851 ## Belongs to the peptidase C14 family. & Contains 2 DED (death effector) domains.
SIZE: 521 amino acids; 58951 Da
SUBUNIT: Heterotetramer that consists of two anti-parallel arranged heterodimers, each one formed by a 23/17 kDa (p23/17) (depending on the splicing events) and a 12 kDa (p12) subunit (By similarity). Self-associates. Interacts with FADD and CASP8. Found in a Fas signaling complex consisting of FAS, FADD, CASP8 and CASP10.
TISSUE SPECIFICITY: Detectable in most tissues. Lowest expression is seen in brain, kidney, prostate, testis and colon.
PTM: Cleavage by granzyme B and autocatalytic activity generate the two active subunits. & Phosphorylated upon DNA damage, probably by ATM or ATR.
DISEASE: SwissProt: Q92851 # Defects in CASP10 are the cause of autoimmune lymphoproliferative syndrome type 2A (ALPS2A) [MIM:603909]. ALPS2 is characterized by abnormal lymphocyte and dendritic cell homeostasis and immune regulatory defects. & Defects in CASP10 are a cause of familial non-Hodgkin lymphoma (NHL) [MIM:605027]. NHL is a cancer that starts in cells of the lymph system, which is part of the body's immune system. NHLs can occur at any age and are often marked by enlarged lymph nodes, fever and weight loss. & Defects in CASP10 are a cause of gastric cancers [MIM:137215].
SIMILARITY: SwissProt: Q92851 ## Belongs to the peptidase C14 family. & Contains 2 DED (death effector) domains.
Components:
- Cell Lysis Buffer: 25 mL
- 2X Reaction Buffer: 2 mL.
- DTT (1 M): 100 μL
- AEVD-AFC Substrate (1 mM): 125 μL
Brand Family:
Chemicon
UniProt Number:
Storage Conditions:
We recommend storing reagents at -20°C upon arrival. (Store Cell Lysis Buffer and 2X Reaction Buffer at 4°C after opening) upon arrival. The performance of this product is guaranteed for 6 months.
Detection Methods:
Fluorescent
Gene Symbol:
- CASP10
- ALPS2
- MCH4
- FLICE2
- CASP-10
- EC 3.4.22.63 [Contains: Caspase-10 subunit p23/17
- Caspase-10 subunit p12].
Product Name:
Caspase 10 Fluorometric Assay Kit, AEVD
Entrez Gene Number:
Materials Required but Not Delivered:
· Microcentrifuge and 1.5 mL Microcentrifuge tubes
· 37°C water bath or incubator
· Fluorometer equipped with a 400-nm excitation filter and 505-nm emission filter.
· 96 well plate
· 37°C water bath or incubator
· Fluorometer equipped with a 400-nm excitation filter and 505-nm emission filter.
· 96 well plate