The mouse testis was immediately dissected and fixed with 2% paraformaldehyde in 0.05% PBST (PBS with 0.05% Triton X-100) at room temperature for 5 min. The fixed sample was placed on a slide glass and squashed by placing a cover slip on top and pressing down. The sample was immediately flash frozen in liquid nitrogen, and the slides were stored at −80°C for further immunofluorescence experiments (Wellard et al., 2018 (link)). After removing the coverslips, the slides were washed with PBS three times and then treated with 0.1% Triton X-100 for 10 min, rinsed three times in PBS, and blocked with 5% bovine serum albumin (Amresco, AP0027). The primary antibody was added to the sections and incubated at 4°C overnight, followed by incubation with the secondary antibody. The nuclei were stained with DAPI. The immunofluorescence images were taken immediately using an LSM 780 microscope (Zeiss) or SP8 microscope (Leica).
Ap0027
Manufactured by Avantor
Sourced in United States
The AP0027 is a laboratory equipment product. It serves as a core function for laboratory applications. No further details can be provided in an unbiased and factual manner.
Automatically generated - may contain errors
3 protocols using ap0027
1
Immunofluorescence Analysis of Mouse Testis
2
Immunofluorescence Analysis of Spermatocytes
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The chromosome spreads of spermatocytes were washed with PBS for 3 times and blocked with 5% bovine serum albumin (AP0027, Amresco, Solon, OH). Primary antibodies were incubated at 4 °C overnight, followed by incubation with the secondary antibodies. The nuclei were stained with 4′, 6-diamidino-2-phenylindole (DAPI). The images were taken immediately using an LSM 780 microscope (Zeiss, Oberkochen, Germany) or a TCS SP8 microscope (Leica, Wetzlar, Germany). 5-μm sections mounted on glass slides were first deparaffinized and then boiled for 15 min in sodium citrate buffer for antigen retrieval. After washing with PBS, sections were blocked and followed by antibody incubation as described above. To detect apoptotic cells in testis, we used the terminal deoxynucleotidyl transferase dUTP nick end-labeling (TUNEL) assay kit (In Situ Cell Death Detection Kit; Roche, 11684795910) according to the manufacturer’s instructions. [53 (link)] Briefly, sections of the testes were deparaffinized and boiled for 15 min in sodium citrate buffer for antigen retrieval. After treated with H2O2 for 10 min at room temperature and sodium citrate for 2 min on ice, the slides were rinsed twice with PBS, the TUNEL reaction mixture was added and incubated in a humidified atmosphere for 60 min at 37 °C in the dark, following by immunofluorescence staining as detailed above.
3
Immunofluorescence Staining of Oocytes
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Oocytes were fixed and permeabilized in 4% PFA and 0.5% Triton X‐100 for 20 min at room temperature. After blocking with 1% bovine serum albumin (BSA, AP0027, Amresco, USA) in PBS, oocytes were incubated with primary antibodies diluted in 1% BSA overnight at 4 °C. Following three washes with PBS, oocytes were labelled with secondary antibodies for 1 h at 37 °C, and then stained with 4′,6‐diamidino‐2‐phenylindole (DAPI) for 5 min. Finally, oocytes were mounted on glass slides and observed with an LSM 780 microscope (Zeiss, Oberkochen, Germany) or SP8 microscope (Leica, Wetzlar, Germany).
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!