Western Blotting and Immunostaining of Meiotic Proteins

Anti-myc (9E10, sc-40, Santa Cruz), anti-tubulin (MCA77G, Bio-Rad/Serotec, Ltd), anti-Cdc5 (sc-33625, Santa Cruz), and histone H3 phospho-T11 (ab5168, Abcam) were used for western blotting. Rabbit anti-Rad51^{21 (link)} and guinea pig anti-Rad51^{73 (link)}, rabbit anti-Hop1^{74 (link)} and rabbit anti-Rfa2^{75 (link)} were home-made and used for staining. Rabbit anti-Ndt80 was a gift by M. Lichten. The secondary antibodies for immuno-staining were Alexa Fluor 488 (Goat, A-21206, Thermo Fisher Scientific) and 594 (Goat, A-211012, Thermo Fisher Scientific) IgG used at a 1/2000 dilution. For western blotting, alkaline phosphatase-conjugated secondary antibodies (Promega) were used.

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Sampathkumar A., Zhong C., Tang Y., Fujita Y., Ito M, & Shinohara A. (2024). Replication protein-A, RPA, plays a pivotal role in the maintenance of recombination checkpoint in yeast meiosis. Scientific Reports, 14, 9550.

Publication 2024

ab5168, Alexa Fluor 488 alkaline phosphatase-conjugated secondary antibodies

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Variable analysis

independent variables

Anti-myc (9E10, sc-40, Santa Cruz)
Anti-tubulin (MCA77G, Bio-Rad/Serotec, Ltd)
Anti-Cdc5 (sc-33625, Santa Cruz)
Histone H3 phospho-T11 (ab5168, Abcam)
Rabbit anti-Rad51 (21)
Guinea pig anti-Rad51 (73)
Rabbit anti-Hop1 (74)
Rabbit anti-Rfa2 (75)
Rabbit anti-Ndt80

dependent variables

Western blotting

control variables

Secondary antibodies for immunostaining (Alexa Fluor 488 and 594, Thermo Fisher Scientific)
Alkaline phosphatase-conjugated secondary antibodies for western blotting (Promega)

positive controls

Rabbit anti-Rad51 (21)
Guinea pig anti-Rad51 (73)
Rabbit anti-Hop1 (74)
Rabbit anti-Rfa2 (75)

negative controls

Not explicitly mentioned

Annotations

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