Cells were grown on glass coverslips. After at least 24 h of culture, cells were fixed with 4% paraformaldehyde, permeabilized with 0.5% Triton X-100 and stained with primary antibodies over-night at 4 °C. 53BP1 (NB100-304) and FANCD2 (NB100-182) antibodies were purchased from Novus Biologicals, γH2AX antibody from Merck/Millipore (05–636), and RAD51 antibody from Santa Cruz biotechnology (sc-8349). Cells were washed three times with PBS Tween 0.1% and incubated with the secondary antibodies for 1 h (Rhodamine Red X (R6394) and AlexaFluor 488 Goat anti-mouse (A11017) or anti-rabbit (A11070), purchased from Invitrogen). DNA was stained with 4.6-diamino-2-phenyl indole (DAPI). Cells were counted positive for foci formation when >10 foci/nuclei were detected.
53bp1 nb100 304 and fancd2 nb100 182
Manufactured by Novus Biologicals
53BP1 (NB100-304) is a protein involved in the DNA damage response pathway. It functions as a key regulator of DNA double-strand break repair. FANCD2 (NB100-182) is a protein that plays a critical role in the Fanconi anemia DNA repair pathway, which is responsible for the repair of DNA interstrand crosslinks.
Automatically generated - may contain errors
2 protocols using 53bp1 nb100 304 and fancd2 nb100 182
1
Immunofluorescence Staining of DNA Damage Markers
2
Immunofluorescence Assay for DNA Damage
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were grown on glass coverslips. After at least 24 hours of culture, cells were fixed with 4% paraformaldehyde, permeabilized with 0.5% Triton X-100 for 15 minutes, blocked in 3% BSA and stained with primary antibodies (dilution 1:3000) overnight at 4 °C in blocking solution. For FANCD2 detection, a pre-extraction step (0.5% Triton X-100 for 5 minutes) was performed before fixation. 53BP1 (NB100-304) and FANCD2 (NB100-182) antibodies were purchased from Novus Biologicals and γH2AX antibody from Merck/Millipore (05-636). Cells were washed three times with PBS Tween 0.1% and incubated with the secondary antibodies (dilution 1:800) for 1 hour (Rhodamine Red X (R6394) and AlexaFluor 488 Goat anti-mouse (A11017) or anti-rabbit (A11070), purchased from Invitrogen). DNA was stained with DAPI. Cells were counted positive for γH2AX and 53BP1 foci formation when > 10 foci/nuclei were detected and counted positive for FANCD2 when > 3 foci/nuclei were detected.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!