The fresh samples of testes from newborn Shaziling boars were fixed in 4% of formaldehyde for 72 h. Next, testes sections were made using routine histological methods. Slides were incubated for two hours, at 37 °C, with anti-SOX9 (1:3000, Proteintech Group, Chicago, IL, USA, 67439-1-Ig) or/and anti-FZD7 (1:1000, Bioss, Beijing, China, bs-5125R). After incubating with secondary antibodies for 1 h, at 37 °C, sections were soaked three times in phosphate-buffered saline (PBS) for 5 min each. DAPI (50 ng/mL, Sigma, Louis, MO, USA) was dripped onto testes slices and incubated at 26 °C, for 5 min, and then washed three times with PBS. All images were shot using a confocal laser scanning microscope (Zeiss LSM 510 META, GER).
Anti sox9
Manufactured by Proteintech
Sourced in United States
Anti-SOX9 is a primary antibody used to detect the transcription factor SOX9 in various applications. SOX9 is a key regulator of chondrocyte differentiation and is involved in the development of various tissues including cartilage, bone, and the male reproductive system.
Automatically generated - may contain errors
Lab products found in correlation
Lsm 510 meta, by Zeiss (2 mentions)
4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions)
Dapi solution, by Merck Group (1 mentions)
Alexa fluor 488 donkey anti rabbit igg h l, by Thermo Fisher Scientific (1 mentions)
Dapi solution, by Thermo Fisher Scientific (1 mentions)
Anti vimentin, by Abcam (1 mentions)
Anti ki67, by Abcam (1 mentions)
Prolong diamond antifade mountant, by Thermo Fisher Scientific (1 mentions)
Anti eed, by Cell Signaling Technology (1 mentions)
Anti runx2, by Cell Signaling Technology (1 mentions)
Anti sox10, by Cell Signaling Technology (1 mentions)
Anti col1a2, by Proteintech (1 mentions)
Anti brdu, by Abcam (1 mentions)
3 protocols using anti sox9
1
Immunohistochemical Analysis of Testicular Development
2
Immunohistochemical Analysis of Boar Testes
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Testicular samples, freshly collected from 23-day-old Shaziling boars, were promptly fixed in 4% formaldehyde for 72 h. Sections of these samples were then prepared following standard histological protocols. The slides were incubated for 2 hours at 37°C with either anti-SOX9 (1:3000, Proteintech Group, Chicago, IL, USA, 67439-1-Ig) or anti-DNMT3A (1:200, HuaBio, Hangzhou, China, ST04-78). After an additional hour of incubation at 37°C with secondary antibodies, the sections were rinsed in phosphate-buffered saline (PBS) thrice, for 5 min each time. The testes slices were then treated with DAPI solution (50 ng/mL, Sigma, Louis, MO, USA), and incubated at 26°C for 5 min, followed by another three rinses with PBS. The final images were captured with a Zeiss LSM 510 META confocal laser scanning microscope.
3
Immunofluorescence Staining of Cryosections and Primary Cells
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Immunofluorescence of cryosections was performed on glass slides. Immunofluorescence for primary cells was performed on glass cover slips. Primary cells were fixed in 4% PFA for 10 minutes at room temperature. Antibody incubations were performed in blocking solution (0.1% Triton X-100, 1% BSA, 10% donkey serum) for 2hr at room temperature or overnight at 4°C. Slides or cover slips were mounted in ProLong Diamond Antifade Mountant (Thermo Scientific, cat# P36965). Primary antibodies used in this study were anti-Sox9 (1:500, Proteintech, cat# 67439-1-Ig), anti-Runx2 (1:500, Cell Signaling, cat# D1L7F), anti-Ki67 (1:2000, Abcam, cat# ab15580), anti-Vimentin (1:1000, Abcam, cat# ab8069), anti-BrdU (1:1000, Abcam, cat# ab6326), anti-Sox10 (1:500, Cell Signaling, cat# D5V9L), anti-ALPL (1:1000, Invitrogen, cat# PA5-47419), anti-Col1a2 (1:500, Proteintech, cat# 66761-1-Ig), anti-Sox6 (1:1000, Proteintech, cat# 14010-1-AP), and anti-Eed (1:1000, Cell Signaling, cat# E4L6E). Secondary antibodies used in this study were Donkey anti-Rabbit IgG (H+L) Alexa Fluor Plus 555 (1:1000, Thermo Scientific, cat# A32794), Donkey anti-Rabbit IgG (H+L) Alexa Fluor 488 (1:1000, Thermo Scientific, cat# A21206), and Donkey anti-Rabbit IgG (H+L) Alexa Fluor Plus 568 (1:1000, Thermo Scientific, cat# A10042). DAPI solution (1:5000, Thermo Scientific, cat# 62248) was added to secondary antibody solutions.
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