Immunostaining of Meiotic Chromosome Proteins

Fetal oocyte cytospreads were performed as previously described [49 (link)]. Slides were incubated with the first antibody against rabbit/mouse SCP3 (rabbit polyclonal, Novus Biologicals Littleton, NB300-232, USA; or mouse polyclonal, Abcam, ab97672, USA), γH2AX (mouse polyclonal, Abcam, ab26350) or RAD51 (rabbit polyclonal, Abcam, ab133534), for 8 h at 37 °C. After overnight blocking with TBS supplemented with 10% goat serum (Boster, AR009, Wuhan, China), Cy3-labeled goat anti-rabbit (Beyotime, A0516) were used to label SCP3/RAD51 and FITC-conjugated goat anti-mouse antibodies (Beyotime, A0568), were used to label γH2AX/SCP3 at 1:100 dilution for 30 min at 37 °C in the dark; DNA was stained with Hoechst 33342. Pictures were taken with an Olympus fluorescence microscope (BX51).

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Zhai Q.Y., Ge W., Wang J.J., Sun X.F., Ma J.M., Liu J.C., Zhao Y., Feng Y.Z., Dyce P.W., De Felici M, & Shen W. (2018). Exposure to Zinc oxide nanoparticles during pregnancy induces oocyte DNA damage and affects ovarian reserve of mouse offspring. Aging (Albany NY), 10(8), 2170-2189.

Publication 2018

NB300-232 ab97672 ab26350 ab133534 AR009 Cy3-labeled goat anti-rabbit A0516 A0568

Anti antibodies Antibody Biologicals Dilution Fetal Fitc Fluorescence microscope Goat Hoechst 33342 Mouse Novus Oocyte Rabbit Serum

Corresponding Organization :

Other organizations : Qingdao Agricultural University, Heilongjiang Academy of Sciences, Institute of Animal Sciences, Auburn University, University of Rome Tor Vergata

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Variable analysis

independent variables

First antibody against rabbit/mouse SCP3
First antibody against γH2AX
First antibody against RAD51

dependent variables

Localization and expression of SCP3
Localization and expression of γH2AX
Localization and expression of RAD51

control variables

Incubation time of 8 h at 37 °C for the first antibodies
Overnight blocking with TBS supplemented with 10% goat serum
Incubation time of 30 min at 37 °C for the secondary antibodies
DNA staining with Hoechst 33342

positive controls

Not mentioned

negative controls

Not mentioned

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