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Anti nobox

Manufactured by Abcam

Anti-Nobox is a protein-specific antibody that recognizes the Nobox (NOBOX) protein. Nobox is a homeobox-containing transcription factor involved in oocyte development and early embryonic development. The Anti-Nobox antibody can be used for the detection and analysis of the Nobox protein in various applications, such as Western blotting, immunohistochemistry, and immunofluorescence.

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2 protocols using anti nobox

1

Oocyte Maturation Analysis via Microscopy

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To detect oocyte maturation, ovaries were fixed and embedded in paraffin, and serial sections (5-µm) were stained with hematoxylin and eosin solutions (Sigma). Images were detected using a Zeiss Axioskop2 MAT microscope (Carl Zeiss MicroImaging), and the total number of follicles was determined. For immunofluorescence analysis, ovary tissues were embedding in a cryomold with compound (Sakura), frozen on dry ice and stored at −80 °C. For imaging, the samples were serially sectioned (5-µm) and fixed with 100% methanol. The sections were washed, blocked for 1 h at RT using Protein Block Serum-Free buffer, and then incubated overnight at 4 °C with a 1:100 dilution of primary antibodies as follows: anti-Nanos3 (Abcam), anti-Lhx8 (Santa Cruz Biotechnology), anti-Nobox (Abcam) and anti-stem121 (StemCells, Inc.) in Antibody Diluent with Background Reducing Components (Dako). After then, the sections were incubated for 90 min at RT with 1:200 dilutions of Alexa Fluor 594-conjugated goat anti-rabbit IgG and Alexa Fluor 488-conjugated mouse anti-goat IgG (Santa Cruz Biotechnology). The sections were counterstained with 4′,6-diamidino-2-phenylindole (DAPI, Dako). The number of follicles was determined, and follicles less than 100 µm in size in each rat were quantified. All experiments were performed in triplicate.
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2

Western Blot Analysis of Ovarian Markers

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Ovary tissues in each group were homogenized and lysed in protein lysis buffer (Sigma). Equal amounts of protein lysates from individual animals were pooled, resolved on 10% sodium dodecyl sulfate polyacrylamide gels, and transferred to PVDF membranes (Bio-Rad Laboratories). The membranes were blocked and incubated with primary antibodies of anti-Lhx8 (1:1,000, Santa Cruz Biotechnology), anti-Nanos3 (1:1,000, Abcam, Cambridge) and anti-Nobox (1:1,000, Abcam) and reacted with a secondary antibodies of anti-rabbit IgG (1:10,000; Bio-Rad Laboratories) or anti-goat IgG (1:5,000; Santa Cruz Biotechnology). The bands were detected using a ChemiDoc system (Bio-Rad Laboratories). All reactions were performed in triplicate for quantification.
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