Apoptosis Resource: Changes in the Nucleus
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Measurement of Apoptosis-Induced Changes in the Nucleus
DNA fragmentation is one of the hallmarks of apoptosis. DNA is first cleaved into large fragments (50-200 kb), followed by cleavage into smaller fragments called nucleosomal units (180-200 bp). The primary DNase responsible for this DNA cleavage is CAD (caspase-activated DNase) or DFF40, (DNA fragmentation factor). CAD is maintained in an inactive state by forming a complex with ICAD (inhibitor of CAD). Upon receiving an appropriate apoptotic signal, caspase-3 cleaves ICAD, and CAD is released to cleave chromosomal DNA. CAD contains a nuclear localization signal, which allows it to cleave only DNA in the nucleus.
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 FragEL™ DNA Fragmentation Detection Kit, Fluorescent-TdT Enzyme Cat. No. QIA39
This kit is a fluorescein-conjugated version of the TdT Colorimetric FragEL™ DNA fragmentation Detection Kit (Cat. No. QIA33). Terminal deoxynucleotidyl transferase (TdT) binds to exposed 3‘-OH ends of DNA fragments generated in response to apoptotic events, and catalyzes the addition of fluorescein-labeled and unlabeled deoxynucleotides. When excited, fluorescein generates an intense signal that can be detected either by fluorescence microscopy or by flow cytometry. The mounting medium sustains the fluorescent signal from samples labeled on slides and aids in the morphological evaluation and characterization of normal and apoptotic cells. Non-apoptotic cells can be visualized using a DAPI filter. |
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| Use of Agarose Gel Electrophoresis to Measure DNA Fragmentation |
Apoptotic nucleosomal fragments (180-200 bp) can be resolved by agarose gel electrophoresis to detect DNA ladders. DNA laddering may not be detectable in all cells undergoing apoptosis (e.g., nerve cells, hepatocytes, and embryonal-fibroblasts), or when the number of cells or the sample size is limited.
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| Measurement of Changes in the Nucleus: Agarose Gel Electrophoresis to Measure DNA Fragmentation |
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| Suicide Track™ DNA Ladder Isolation Kit |
Adherent or suspension cells |
25 Extractions |
AM41 |
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| Measurement of Changes in the Nucleus: Use of Vital Dyes |
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| Acridine Orange |
113000 |
| Bisbenzimide H 33258 Fluorochrome, Trihydrochloride |
382061 |
| Bisbenzimide H 33342 Fluorochrome, Trihydrochloride |
382065 |
| DAPI, Dihydrochloride |
268298 |
| Propidium Iodide |
537059 |
| Propidium Iodide Solution |
537060 |
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| Use of Terminal Deoxyuridine Nucleotide End Labeling (TUNEL) to Measure DNA Fragmentation |
| DNases generate free 3´-hydroxyl termini, which in turn can be labeled with bromolated deoxyuridine triphosphate nucleotides (Br-dUTP). The reaction is catalyzed by deoxynucleodidyl transferase (TdT) and Br-dUTP sites can be detected with a BrdU antibody conjugated to a fluorophore. Since non-apoptotic cells lack exposed 3´-hydroxyl ends, little or no Br-dUTP is incorporated. |
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| Use of TUNEL to Measure DNA Fragmentation: DNA Fragmentation Based Kits |
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| Apo-BrdU™ Kit |
FC, IF |
Cells |
60 Tests |
CBA040 |
| Apo-Direct™ Kit |
FC |
Cells |
50 Tests |
CBA041 |
| FragEL™ DNA Fragmentation Detection Kit, Colorimetric - Klenow Enzyme |
FS, PS |
Frozen or paraffin sections or fixed-cell preparations |
50 Tests |
QIA21 |
| FragEL™ DNA Fragmentation Detection Kit, Colorimetric - TdT Enzyme |
FS, PS |
Frozen or paraffin sections or fixed-cell preparations |
50 Tests |
QIA33 |
| FragEL™ DNA Fragmentation Detection Kit, Fluorescent - TdT Enzyme |
FC |
Frozen or paraffin sections, fixed cell preparations or cell suspensions |
50 Tests |
QIA39 |
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| Use of TUNEL to Measure DNA Fragmentation: Other Related Kits |
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| Cell Death Detection (Nuclear Matrix Protein) ELISA Kit |
ELISA |
Tissue culture supernatant |
96 Tests |
QIA20 |
| Nucleosome ELISA Kit |
ELISA |
Cell lysates |
96 Tests |
QIA25 |
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