Cell death occurs by two major mechanisms, necrosis and apoptosis. Apoptsis is also known as programmed cell death or ankoikis (a form of apoptosis which is induced by anchorage-dependent cells detaching from the surrounding extracellular matrix). Apoptosis leads to the elimination of cells without releasing harmful substances into the surrounding area. Too little or too much apoptosis plays a role in a great many diseases. When apoptosis functions inappropriately, cells that should be eliminated survive and potentially become immortal, as in cancer or leukemia. When apoptosis works overly well, too many cells may ÔdieÕ and the result may be grave tissue damage. This is the case in stroke and neurodegenerative disorders such as Alzheimer, Huntington and Parkinson diseases. The term ÔapoptosisÕ refers only to the structural changes a cell goes through during the process of programmed cell death and not to the process itself. Classical necrotic cell death occurs due to noxious injury or trauma to the cell while apoptosis is an energy dependant mechanism that takes place during normal cell development. While necrotic cell death results in cell lysis, cellular apoptosis is characterized morphologically by cell shrinkage, nuclear pyknosis, chromatin condensation, and blebbing of the plasma membrane. Apoptosis is the result of a cascade of molecular and biochemical events involving endogenous endonucleases that cleave DNA into the prototypical Ôladder of DNA fragmentsÕ that may be visualized in agarose gels. Observation of oligonucleosomal DNA fragments by DNA laddering has long been the most acceptable and only available assay for the detection of apoptosis.
The ApoMark™ FL kit supplies sufficient reagents to stain 30 or 60 specimens of approximate size 2.5 cm2. Components shown below are for 30 specimen kit: Component 1: Proteinase K, pH-stabilized Solution (50μl) Component 2: TdT Equilibration Buffer (4 ml) Component 3: TdT labeling reaction mix - Optimized mix of labeled and unlabeled nucleotides (1.3 ml) Component 4: TdT enzyme: Terminal Deoxynucleotidyl Transferase (40 μl) Component 5: Stop Buffer (4 ml) Component 6: Blocking Buffer (12 ml) Component 7: 25X Conjugate: SA-Fluorochrome (150μl)
Antigen-antibody binding interaction
ApoMark Apoptosis Kit FL488 (30 Tests)
Antibody's suited for
ApopMark™ Apoptosis Detection Kit allows the recognition of apoptotic nuclei in paraffin-embedded tissue sections, frozen tissue section, or in preparations of single cell suspensions fixed on slides. In this assay Terminal deoxynucleotidyl Transferase (TdT) binds to exposed 3’-OH ends of DNA fragments generated in response to apoptotic signals and catalyzes the addition of biotin-labeled deoxynucleotides. Biotinylated nucleotides are detected using a streptavidinfluorochrome conjugate which reacts with biotin.
ApoMark™ FL kit components are shipped on cold pack. Upon receipt, store kit at -20°C in a non-frost-free freezer. Thirty (30) minutes prior to use of each component, thaw component and keep on cold block or on ice. Return to -20°C immediately after use. Special care should be taken to keep TdT (component 4), Labeling Reaction Mix (component 3) and 25x Conjugate (component 7) cold.
ApoMark Apoptosis Kit FL488 (30 )
ApoMark programmed cellular destruction reagent FL488 (30 Tests)