The ovarian tissues were cut into 5-μm-thick sections. After antigen retrieval, the ovarian sections were incubated overnight with a rabbit polyclonal antibody against AMH (ab103233, 1:2000, Abcam). After washing, the immunoperoxidase-labeled Donkey Anti-Rabbit IgG secondary antibody (1:2000, LS-C60950, LSBio) was applied to all slides at room temperature for 1 h. After washing with PBS three times, DBA was used for color rendering, and the nuclei were re-stained with hematoxylin for 2 min. Finally, the slides were sealed with neutral adhesive for observation, and the AMH signals were acquired using a light microscope (BX51, Olympus, Tokyo, Japan).
Ab103233
Manufactured by Abcam
Sourced in China, United Kingdom, United States
Ab103233 is a primary antibody that targets a specific protein. It is intended for use in various laboratory applications, such as immunoassays and immunohistochemistry. The product is supplied as a liquid solution.
Automatically generated - may contain errors
Lab products found in correlation
Alexa fluor 594, by Proteintech (1 mentions)
Af5366, by Affinity Biosciences (1 mentions)
Tcs sp5 confocal microscope, by Leica (1 mentions)
Citrate antigen retrieval solution, by Beyotime (1 mentions)
Ab6721, by Abcam (1 mentions)
Sc 47778, by Santa Cruz Biotechnology (1 mentions)
Clarity western ecl substrate, by Bio-Rad (1 mentions)
4 protocols using ab103233
1
Immunohistochemical Analysis of AMH
2
AMH Expression and Localization by IF
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KGN cells seeded onto glass coverslips in six-well plates at a density of 5 × 104 were transfected with plasmids for expression of FLAG-tagged AMH and treated with CTX, GnRHa, or CTX and GnRHa. After 48 h, immunofluorescence staining assays were carried out, as previously described39 (link), with primary antibodies for AMH (ab103233, Abcam, Shanghai, China) and GRP78 (AF5366, Affinity, Jiangsu, China), and secondary goat anti-rabbit antibodies conjugated to FITC or Alexa Fluor 594 (Proteintech). Images were visualized and captured with a TCS SP5 confocal microscope Leica (Shanghai, China).
3
Immunocytochemistry Quantification of AMH Expression
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Immunocytochemistry was performed according to routine immunohistochemistry methodology. In brief, KGN cells fixed in PFA were paraffin-embedded and serially sectioned (5-mm sections). Then, these sections were deparaffinized, and endogenous peroxidase was inactivated by treatment with 3% (w/v) hydrogen peroxide. Citrate antigen retrieval solution (Beyotime, Shanghai, China) was used for antigen retrieval for 30 mins at 95–100 °C. Slide-mounted cell sections were incubated with an AMH antibody (ab103233, Abcam, Cambridge, UK) overnight at 4 °C and an HRP-conjugated secondary antibody for 1 h at room temperature. A GTVision III detection System/Mo&Rb kit (DAKO, K5007, Denmark) was used for subsequent visualization steps. Cell sections were stained with diaminobenzidine substrate, and nuclei were counterstained with hematoxylin. Light microscopy was used for image visualization and capture. Result quantified by image analysis software (ImageJ 1.51e, NIH Image).
4
Quantification of Ovarian AMH Levels
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Western blot analyses for anti-Müllerian hormone (AMH) production were performed on human ovarian tissues. Western blot analyses were performed as previously described [34 (link)]. Proteins were detected by incubation with anti-AMH (1:100 dilution, catalogue #ab103233; Abcam), and anti-β-actin (1:1000, catalogue #sc-47778; Santa Cruz Biotechnology, Santa Cruz, CA, USA) primary antibodies at 4 °C overnight with gentle shaking. Following incubation, the proteins incubated with goat anti-rabbit secondary antibody (1:5000 dilution, catalogue #ab6721; Abcam) in 1× tris-buffered saline (TBS) containing 3% skim milk and 0.05% Tween-20 at room temperature for 90 min. Immunoreactive proteins were visualized by chemiluminescence using Clarity Western ECL Substrate (catalogue #1705060, Bio-Rad Laboratories) and detected on medical X-ray blue film (Agfa-Gevaert, Mortsel, Belgium). Western blotting was repeated five times for AMH analyses. The band intensities were quantified relative to intensity of beta-actin by densitometry using imageJ software.
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