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. Author manuscript; available in PMC: 2012 Jan 1.
Published in final edited form as: Methods Mol Biol. 2011;780:491–516. doi: 10.1007/978-1-61779-270-0_29

Figure 7.

Figure 7

Primary and Secondary Antibody Optimization, Optimization of PARP1 Amount

(a) Plate setup and absorbance readings for primary and secondary antibody optimization and optimization of PARP1 amount. 10H primary antibody dilutions: columns 1, 2, 7, 8: 1:1000; columns 3, 4, 9, 10: 1:2000; columns 5, 6, 11, 12: 1:4000. Amount of PARP used in reaction: columns 1-6: 0.15 u and columns 7 -12: 0.075U. Secondary antibody dilutions: rows A-D: 1:1500; rows F-H: 1:3000.

(b) Primary and secondary antibody dilutions optimization. Amount of PARP1 in reaction. Secondary antibody dilution 1:3000 reduces sensitivity of reaction, while for primary antibody, a dilution of 1:4000 will also give a good DMSO/3AB signal ratio. Decreasing the amount of PARP1 to 0.075 U/well also does not significantly change DMSO/3AB signal ratio.