Skip to Content
MilliporeSigma
All Photos(1)

Key Documents

APT423

Sigma-Aldrich

CaspaTag Caspase 3,7 In Situ Assay Kit, Fluorescein

The CaspaTag Caspase-3/7 In Situ Assay Kit, Fluorescein for Flow Cytometry is a fluorescent-based assay for detection of active caspase-3 or caspase-7 in cells undergoing apoptosis.

Sign Into View Organizational & Contract Pricing


About This Item

UNSPSC Code:
12161503
eCl@ss:
32161000
NACRES:
NA.54

Quality Level

species reactivity (predicted by homology)

mammals

manufacturer/tradename

CaspaTag
Chemicon®

technique(s)

activity assay: suitable
flow cytometry: suitable

NCBI accession no.

UniProt accession no.

detection method

fluorometric

shipped in

wet ice

Gene Information

human ... CASP3(836)

General description

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 cell1.

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 tetramer2-4. In common with other proteases, caspases are synthesized as precursors that undergo proteolytic maturation, either autocatalytically or in a cascade by enzymes with similar specificity5.

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 residue6. 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.

Application

Research Category
Apoptosis & Cancer
The CHEMICON® CaspaTag Caspase-3/7 In Situ Assay Kit, Fluorescein 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.

Test Principle

CHEMICON′s CaspaTag 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 caspase7. 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.
The CaspaTag Caspase-3/7 In Situ Assay Kit, Fluorescein for Flow Cytometry is a fluorescent-based assay for detection of active caspase-3 or caspase-7 in cells undergoing apoptosis.

Components

FLICA Reagent (FAM-DEVD-FMK): One lyophilized vial

10X Wash Buffer: 15 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

Storage and Stability

· Store unopened kit materials at 2-8°C up to their expiration date.

· Reconstituted FLICA Reagent (150X) should be frozen at -20°C 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 -20°C for 2 weeks.

Legal Information

CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany

Disclaimer

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.

Pictograms

Skull and crossbonesHealth hazard

Signal Word

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

Target Organs

Eyes,Central nervous system, Respiratory system

Storage Class Code

6.1C - Combustible acute toxic Cat.3 / toxic compounds or compounds which causing chronic effects


Certificates of Analysis (COA)

Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.

Already Own This Product?

Find documentation for the products that you have recently purchased in the Document Library.

Visit the Document Library

Kazunori Okano et al.
Biochemistry and biophysics reports, 24, 100818-100818 (2020-10-22)
The techniques for inducing the death of specific cells in tissue has attracted attention as new methodologies for studying cell function and tissue regeneration. In this study, we show that a sequential process of targeted cell death and removal can
Stefan Frantz et al.
European heart journal, 34(16), 1233-1244 (2011-12-27)
Cardiac hypertrophy is a common and often lethal complication of arterial hypertension. Elevation of myocyte cyclic GMP levels by local actions of endogenous atrial natriuretic peptide (ANP) and C-type natriuretic peptide (CNP) or by pharmacological inhibition of phosphodiesterase-5 was shown
Hideo A Baba et al.
Liver international : official journal of the International Association for the Study of the Liver, 29(4), 585-592 (2008-11-21)
Survivin regulates cell division and inhibits apoptosis. Liver regeneration is a complex process involving both proliferation and apoptosis. The role of survivin is not well elucidated and no data exist in humans. Seventy per cent liver resection was used to
Christina L Kaiser et al.
Hearing research, 240(1-2), 1-11 (2008-05-20)
Aminoglycoside antibiotics induce caspase-dependent apoptotic death in cochlear hair cells. Apoptosis, a regulated form of cell death, can be induced by many stressors, which activate signaling pathways that result in the controlled dismantling of the affected cell. The caspase family
Yidi Qu et al.
Aging, 13(8), 11150-11169 (2021-04-06)
Alzheimer's disease (AD) is characterized by cognitive decline due to the accumulation of extracellular β-amyloid (Aβ) plaques and neurofibrillary tangles in the brain, which impair glutamate (Glu) metabolism. Deproteinized Calf Blood Extractive Injection (DCBEI) is a biopharmaceutical that contains 17

Our team of scientists has experience in all areas of research including Life Science, Material Science, Chemical Synthesis, Chromatography, Analytical and many others.

Contact Technical Service