The antigen-antibody complexes were visualized by enhanced chemiluminescence ECL detection system; Amersham. Br J Cancer. Bioluminescence Methods and Protocols. Organic Contaminant Detection and Biodegradation Characteristics. Immunoprecipitation and immunoblot analysis.
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Luminescence-Based Cell Viability Testing.
By contrast, IR-induced apoptosis involves activation of a CrmA-insensitive pathway Human cells contain a DNA-activated protein kinase that phosphorylates simian virus 40 T antigen, mouse p53 and the human Ku autoantigen. Overexposed gels demonstrate a minor cleaved fragment of kDa data not shown. In this context, cytochrome c release activates caspase 9, and this event is upstream to activation of caspase 3 The resulting immune complexes were washed three times with lysis buffer, separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis SDS-PAGEand transferred to nitrocellulose filters.
Bioluminescence Methods and Protocols SpringerLink
The contributors-all hands-on researchers who have perfected their protcols-illustrate these powerful methods with a wide variety of applications ranging from the analytical determination titer of specific molecules to the cellular detection of a wide range of pollutants, to microbiological detection and analysis, and the molecular biological manipulation of lux, luc, and gfp structural genes.
|Binding of Ku and c-Abl at the kinase homology region of DNA-dependent protein kinase catalytic subunit. J Cell Biol. Double strand break repair.
Tewari M, Dixit V M. Preimmune rabbit serum PIRS was used as a negative control.
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They were washed with PBS, resuspended in 0. D U cells were treated with 20 Gy of IR and harvested at the indicated times. To address this issue, we first incubated purified DNA-PK—Ku complexes with in vitro-translated caspase 3 to generate its fragments. Front Matter Pages Step-by-step methods for the construction of other recombinant organisms are also included.
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|Weaver D T. As controls, cells were transfected with GFP-expressing empty vector. D U cells were treated with 20 Gy of IR and harvested at the indicated times.
After 12 to 18 h, the cells were harvested and sorted by FACscan analysis. PIRS was used as a negative control of immunoprecipitation. Advertisement Hide.
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Phagocyte Chemiluminescence. Arrows indicate apoptotic cells. To address this issue, we first incubated purified DNA-PK—Ku complexes with in vitro-translated caspase 3 to generate its fragments.
Whole-cell lysate Lysate was used as a positive control for the immunoblot analysis. Editors and affiliations. Scid cells are deficient in Ku and replication protein A phosphorylation by the DNA-dependent protein kinase. The kDa fragment can also undergo further cleavage to a kDa protein that retains the kinase domain