Protocol detail

Multiparametric Imaging of DNA Damage

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IF-FISH was performed as previously described [50 (link), 58 (link)].
Fluorescent probe is Cy3-(TTAGGG)₃ (Panagene). Primary antibodies include anti-53BP1 (Novus Biologicals), anti-RPA2 (EMD Millipore), anti-PCNA (Genetex), anti-PML (Santa Cruz), anti-SMARCAL1 (Santa Cruz), anti-SLX4 (Novus Biologicals), anti-pDNA-PKcs (S2056) (Abcam) and anti-Rad51 (Santa Cruz). Secondary antibodies include DyLight488 conjugated anti-rabbit (Multisciences), DyLight488 conjugated anti-mouse (Multisciences).

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Zhang T., Zhang Z., Shengzhao G., Li X., Liu H, & Zhao Y. (2019). Strand break-induced replication fork collapse leads to C-circles, C-overhangs and telomeric recombination. PLoS Genetics, 15(2), e1007925.

Publication 2019

Cy3-(TTAGGG)3 anti-53BP1 anti-PCNA anti-PML anti-SMARCAL1 anti-SLX4 anti-Rad51

53bp1 Antibodies Biologicals Fish Fluorescent probe Mouse Novus Pcna Pkcs Rabbit Rpa2

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Variable analysis

independent variables

IF-FISH protocol (as previously described in references [50] and [58])
Fluorescent probe (Cy3-(TTAGGG)₃)
Primary antibodies (anti-53BP1, anti-RPA2, anti-PCNA, anti-PML, anti-SMARCAL1, anti-SLX4, anti-pDNA-PKcs (S2056), anti-Rad51)
Secondary antibodies (DyLight488 conjugated anti-rabbit, DyLight488 conjugated anti-mouse)

dependent variables

Experimental observations and measurements related to the cellular localization and distribution of the target proteins

control variables

Not explicitly mentioned

controls

Positive controls: Not specified
Negative controls: Not specified

Annotations

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