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DNA Damage and Repair - Quantification of Sub-Cellular Events Using Automated Confocal Imaging

ssay protocol using only 2 primary antibodies, 1 dye and a commercially available cell line allow this assay to be used in automated high-content screening applications where hands-off operation is required. The BD Pathway Bioimager with its confocal capability, white-light illumination and choice of 16 excitation filters allows development of multi-color assays and would even accommodate a 4th imaging channel. The complexity of change in the cellular phenotype in this assay mandate sophisticated and specialized image and data analysis algorithms. The combination of BD IPLab for Pathway and BD Image Data Explorer allows generation and analysis of high-content data in a drug screening environment.

References

1. Friedberg, E.C, Walker, G.C., Siede, et al . 2005 DNA Repair and Mutagenesis, ASM Press; 2nd edition ISBN : 1555813194

2. Kim, J-S, Krasieva, J_S, Kurumizaka, H, et al . 2005. Independent and sequential recruitment of NHEJ and HR factors to DNA damage sites in mammalian cells. JCB , 170, 341-347

3. Paull T.T., Rogakou E.P., Yamazaki V., Kirchgessner C.U., Gellert M., Bonner W.M..2000. A critical role for histone H2AX in recruitment of repair factors to nuclear foci after DNA damage. Curr Biol. 10, 886-95.

4. Schultz, L.B, Chehab, N.H., Malikzay, A., Halazonetis, T.H., 2000. p53 Binding Protein 1 (53BP1) Is an Early Participant in the Cellular Response to DNA Double-Strand Breaks. JCB , 7, 1381-1390

5. Sedelnikova O.A., Horikawa I., Zimonjic D.B., Popescu N.C., Bonner W.M., Barrett J.C. 2004. Senescing human cells and ageing mice accumulate DNA lesions with unrepairable double-strand breaks. Nature Cell Biology. 6, 168-70.


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