To detect the Izumo1 probe binding to the oolemma, purified pentamerized FLAG-tagged Izumo1 was added to unfixed eggs. After 30 minutes at 37 °C, eggs were fixed with 2 % formalin, washed, and stained with anti-Flag-Cy3 antibody diluted 1:500 in M2 media. Izumo1 binding to HEK293T cells was detected using the same method except that protein and antibody were diluted in PBS/2 % BSA. Juno was detected on eggs fixed with 2 % formalin using the anti-Juno/Folr4 monoclonal antibody diluted to 2 μg/mL in M2 media, washed, followed by an anti-rat Alexa Fluor 488. Stained eggs were mounted on a microscope glass in SlowFade with DAPI (Molecular Probes). Samples were analysed with Leica SP5/DM6000 confocal microscope.
Sp5 dm6000 confocal microscope
Manufactured by Leica
The SP5/DM6000 confocal microscope is a high-performance imaging system designed for advanced microscopy applications. It features a modular design that allows for customization to meet specific research requirements. The microscope provides excellent optical performance, enabling users to capture detailed, high-resolution images of a wide range of samples.
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Lab products found in correlation
2 protocols using sp5 dm6000 confocal microscope
1
Izumo1 Binding to Oolemma and Cells
2
Izumo1 Binding to Oolemma and Cells
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To detect the Izumo1 probe binding to the oolemma, purified pentamerized FLAG-tagged Izumo1 was added to unfixed eggs. After 30 minutes at 37 °C, eggs were fixed with 2 % formalin, washed, and stained with anti-Flag-Cy3 antibody diluted 1:500 in M2 media. Izumo1 binding to HEK293T cells was detected using the same method except that protein and antibody were diluted in PBS/2 % BSA. Juno was detected on eggs fixed with 2 % formalin using the anti-Juno/Folr4 monoclonal antibody diluted to 2 μg/mL in M2 media, washed, followed by an anti-rat Alexa Fluor 488. Stained eggs were mounted on a microscope glass in SlowFade with DAPI (Molecular Probes). Samples were analysed with Leica SP5/DM6000 confocal microscope.
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