DNA Damage Foci Quantification

Cells were grown in chamber slides, treated, fixed, immunostained, and analyzed as previously described [60 (link)]. Cells with more than 10 foci were determined as positive. The primary antibodies used were mouse anti-γH2AX (Abcam) at a dilution of 1:2,000, rabbit anti-53BP1 (Cell Signaling) at a dilution of 1:500, and rabbit anti-RPA70 (Cell Signaling) at a dilution of 1:500. Secondary anti-mouse Alexa 555 and anti-rabbit Alexa 488 were from Invitrogen and were used at a dilution 1:1000.

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Gary C., Hajek M., Biktasova A., Bellinger G., Yarbrough W.G, & Issaeva N. (2016). Selective antitumor activity of roscovitine in head and neck cancer. Oncotarget, 7(25), 38598-38611.

Publication 2016

mouse anti-γH2AX rabbit anti-53BP1 anti-mouse Alexa 555

53bp1 Antibodies Cells Dilution Mouse Rabbit

Corresponding Organization :

Other organizations : Yale University, Children's Cancer Institute Australia, Yale Cancer Center

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Variable analysis

independent variables

Treatment

dependent variables

Number of cells with more than 10 foci

control variables

Cells grown in chamber slides
Fixed and immunostained cells
Primary antibody dilutions (mouse anti-γH2AX at 1:2,000, rabbit anti-53BP1 at 1:500, rabbit anti-RPA70 at 1:500)
Secondary antibody dilutions (anti-mouse Alexa 555 and anti-rabbit Alexa 488 at 1:1000)

controls

Positive control: Not specified
Negative control: Not specified

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