APT403
CaspaTag Caspase 3,7 In Situ Assay Kit, Fluorescein
The In Situ Caspase Detection Kit for Flow Cytometry use a novel approach to detect active caspases. The methodology is based on Fluorochrome Inhibitors of Caspases (FLICA).
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모든 사진(1)
About This Item
UNSPSC 코드:
12161503
eCl@ss:
32161000
NACRES:
NA.84
추천 제품
Quality Level
종 반응성(상동성에 의해 예측)
mammals
제조업체/상표
CaspaTag
Chemicon®
기술
activity assay: suitable
flow cytometry: suitable
NCBI 수납 번호
검출 방법
fluorometric
배송 상태
wet ice
유전자 정보
human ... CASP3(836)
일반 설명
Apoptosis is an evolutionarily conserved form of cell suicide, which follows a specialized cellular process. The central component of this process is a cascade of proteolytic enzymes called caspases. These enzymes participate in a series of reactions that are triggered in response to pro-apoptotic signals and result in the cleavage of protein substrates, causing the disassembly of the cell (Slee et al. 1999).
Caspases have been identified in organisms ranging from C. elegans to humans. The mammalian caspases play distinct roles in apoptosis and inflammation. In apoptosis, caspases are responsible for proteolytic cleavages that lead to cell disassembly (effector caspases), and are involved in upstream regulatory events (initiator caspases). An active caspase consists of two large and two small subunits that form two heterodimers which associate in a tetramer (Walker et al. 1994; Wilson et al. 1994; Rotonda et al. 1996). In common with other proteases, caspases are synthesized as precursors that undergo proteolytic maturation, either autocatalytically or in a cascade by enzymes with similar specificity (Kumar 1999).
Caspase enzymes specifically recognize a 4 or 5 amino acid sequence on the target substrate which necessarily includes an aspartic acid residue. This residue is the target for cleavage, which occurs at the carbonyl end of the aspartic acid residue (Thornberry et al. 1997). Caspases can be detected via immunoprecipitation, immunoblotting techniques using caspase specific antibodies, or by employing fluorochrome substrates which become fluorescent upon cleavage by the caspase.
Test Principle:
CHEMICON′s In Situ Caspase Detection Kits use a novel approach to detect active caspases. The methodology is based on Fluorochrome Inhibitors of Caspases (FLICA). The inhibitors are cell permeable and non-cytotoxic. Once inside the cell, the inhibitor binds covalently to the active caspase (Ekert et al. 1999). This kit uses a carboxyfluorescein-labeled fluoromethyl ketone peptide inhibitor of caspase-3 (FAM-DEVD-FMK), which produces a green fluorescence. When added to a population of cells, the FAM-DEVD-FMK probe enters each cell and covalently binds to a reactive cysteine residue that resides on the large subunit of the active caspase heterodimer, thereby inhibiting further enzymatic activity. The bound labeled reagent is retained within the cell, while any unbound reagent will diffuse out of the cell and is washed away. The green fluorescent signal is a direct measure of the amount of active caspase-3 or caspase-7 present in the cell at the time the reagent was added. Cells that contain the bound labeled reagent can be analyzed by 96-well plate-based fluorometry, fluorescence microscopy, or flow cytometry.
Application:
The CHEMICON In Situ FLICA Caspase-3/7 Detection Kit is a fluorescent-based assay for detection of active caspase-3 or caspase-7 in cells undergoing apoptosis. The kit is for research use only. Not for use in diagnostic or therapeutic procedures.
Caspases have been identified in organisms ranging from C. elegans to humans. The mammalian caspases play distinct roles in apoptosis and inflammation. In apoptosis, caspases are responsible for proteolytic cleavages that lead to cell disassembly (effector caspases), and are involved in upstream regulatory events (initiator caspases). An active caspase consists of two large and two small subunits that form two heterodimers which associate in a tetramer (Walker et al. 1994; Wilson et al. 1994; Rotonda et al. 1996). In common with other proteases, caspases are synthesized as precursors that undergo proteolytic maturation, either autocatalytically or in a cascade by enzymes with similar specificity (Kumar 1999).
Caspase enzymes specifically recognize a 4 or 5 amino acid sequence on the target substrate which necessarily includes an aspartic acid residue. This residue is the target for cleavage, which occurs at the carbonyl end of the aspartic acid residue (Thornberry et al. 1997). Caspases can be detected via immunoprecipitation, immunoblotting techniques using caspase specific antibodies, or by employing fluorochrome substrates which become fluorescent upon cleavage by the caspase.
Test Principle:
CHEMICON′s In Situ Caspase Detection Kits use a novel approach to detect active caspases. The methodology is based on Fluorochrome Inhibitors of Caspases (FLICA). The inhibitors are cell permeable and non-cytotoxic. Once inside the cell, the inhibitor binds covalently to the active caspase (Ekert et al. 1999). This kit uses a carboxyfluorescein-labeled fluoromethyl ketone peptide inhibitor of caspase-3 (FAM-DEVD-FMK), which produces a green fluorescence. When added to a population of cells, the FAM-DEVD-FMK probe enters each cell and covalently binds to a reactive cysteine residue that resides on the large subunit of the active caspase heterodimer, thereby inhibiting further enzymatic activity. The bound labeled reagent is retained within the cell, while any unbound reagent will diffuse out of the cell and is washed away. The green fluorescent signal is a direct measure of the amount of active caspase-3 or caspase-7 present in the cell at the time the reagent was added. Cells that contain the bound labeled reagent can be analyzed by 96-well plate-based fluorometry, fluorescence microscopy, or flow cytometry.
Application:
The CHEMICON In Situ FLICA Caspase-3/7 Detection Kit is a fluorescent-based assay for detection of active caspase-3 or caspase-7 in cells undergoing apoptosis. The kit is for research use only. Not for use in diagnostic or therapeutic procedures.
애플리케이션
Research Category
Apoptosis & Cancer
Apoptosis & Cancer
The In Situ Caspase Detection Kit for Flow Cytometry use a novel approach to detect active caspases. The methodology is based on Fluorochrome Inhibitors of Caspases (FLICA).
성분
FLICA Reagent (FAM-DEVD-FMK): Four lyophilized vials
10X Wash Buffer: 60 mL
Fixative: 6 mL
Propidium Iodide: 1 mL at 250 μg/mL, ready-to-use
Hoechst Stain: 1 mL at 200 μg/mL, ready-to-use
10X Wash Buffer: 60 mL
Fixative: 6 mL
Propidium Iodide: 1 mL at 250 μg/mL, ready-to-use
Hoechst Stain: 1 mL at 200 μg/mL, ready-to-use
저장 및 안정성
· Store unopened kit materials at 2-8°C up to their expiration date.
· Reconstituted FLICA Reagent (150X) should be frozen at -20ºC for up to 6 months, and may be thawed twice during this time. Aliquot into separate amber tubes if desired. Protect from light at all times.
· Store diluted (1X) wash buffer up to -20ºC for 2 weeks.
· Reconstituted FLICA Reagent (150X) should be frozen at -20ºC for up to 6 months, and may be thawed twice during this time. Aliquot into separate amber tubes if desired. Protect from light at all times.
· Store diluted (1X) wash buffer up to -20ºC for 2 weeks.
법적 정보
CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany
면책조항
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.
신호어
Danger
Hazard Classifications
Acute Tox. 3 Inhalation - Acute Tox. 4 Dermal - Acute Tox. 4 Oral - Carc. 1B - Eye Irrit. 2 - Muta. 2 - Skin Irrit. 2 - Skin Sens. 1 - STOT SE 2 - STOT SE 3
표적 기관
Eyes,Central nervous system, Respiratory system
Storage Class Code
6.1C - Combustible acute toxic Cat.3 / toxic compounds or compounds which causing chronic effects
시험 성적서(COA)
제품의 로트/배치 번호를 입력하여 시험 성적서(COA)을 검색하십시오. 로트 및 배치 번호는 제품 라벨에 있는 ‘로트’ 또는 ‘배치’라는 용어 뒤에서 찾을 수 있습니다.
Qunfeng Cai et al.
Neurobiology of disease, 45(2), 723-732 (2011-11-03)
Cell-cell junctions and junctions between cells and extracellular matrix are essential for maintenance of the structural and functional integrity of the cochlea, and are also a major target of acoustic trauma. While morphological assessments have revealed adhesion dysfunction in noise-traumatized
Bo Hua Hu et al.
Journal of neuroscience research, 88(8), 1812-1821 (2010-01-22)
Acoustic overstimulation causes apoptotic cell death in the cochlea. This death process is mediated, in part, by the mitochondrial signaling pathway involving Bcl-2 family proteins. Myeloid cell leukemia sequence 1 (Mcl-l) is an antiapoptotic member of the Bcl-2 family. Its
Lisa L Cunningham et al.
The Journal of neuroscience : the official journal of the Society for Neuroscience, 22(19), 8532-8540 (2002-09-28)
Aminoglycoside exposure results in the apoptotic destruction of auditory and vestibular hair cells. This ototoxic hair cell death is prevented by broad-spectrum caspase inhibition. We have used in situ substrate detection, immunohistochemistry, and specific caspase inhibitors to determine which caspases
Mohammad Husein Abnosi et al.
Iranian journal of basic medical sciences, 15(4), 900-906 (2013-03-16)
Arsenic compounds are potent human carcinogen and produce a variety of stress responses in mammalian cells. Recently sodium arsenite has been recommended to be used as anti malignancy drug by American food and drug administration (FDA). In this study, we
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