The collected placental tissues were embedded and sectioned. After dewaxing and rehydration, the sections were treated with 3% H2O2 at 22°C for 15 min and blocked with normal goat serum at 22°C for 15 min. Thereafter, the sections were reacted with 50 μL the primary antibodies against EZH2 (1:50, GTX82503, GeneTex), cytokeratin 7 (CK7, 1:100, ab183344, Abcam), and GADD45A (1:100, PA5-88248, Thermo Fisher Scientific) overnight at 4°C, and then with IgG (1:2,000, ab205718, Abcam) at 37°C for 15 min. After that, 40 μL horseradish peroxidase-labeled streptavidin-working solution was added for 15 min. After color development using 3,3’-diaminobenzidine, the sections were counter-stained with hematoxylin for 30s, dehydrated, and sealed for observation. Positive cells were counted under the microscope (Olympus Optical Co., Ltd., Tokyo, Japan) [25 (link)].
Ab183344
Manufactured by Abcam
Sourced in United Kingdom
Ab183344 is a polyclonal antibody targeting Isocitrate Dehydrogenase 1 (IDH1). It is designed for use in various research applications.
Automatically generated - may contain errors
2 protocols using ab183344
1
Immunohistochemical Analysis of Placental Tissues
2
Immunohistochemical analysis of trophoblast cells
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Formalin-fixed paraffin-embedded tissue samples were obtained and stained with anti-human CK7 antibody (1:500 dilution; ab183344; Abcam, Inc., Cambridge, UK) as the positive control for trophoblast cells. Anti-human SIK3 antibody (1:100 dilution; Dr. provided by Neng-Yao Shih), anti-human CD68 antibody (1:500 dilution; M0814; Agilent Technologies, Inc., Santa Clara, CA, USA), or anti-human CD204 antibody (1:200 dilution; MAB1710; Abnova Corporation) was used to stain the trophoblast cells and macrophages near the decidua. The sections were serially dewaxed, rehydrated, and heated with 10 mM sodium citrate (pH 6.0) for 20 min for antigen retrieval. After endogenous peroxidases were blocked with 3% hydrogen peroxide, the sections were incubated with the primary antibody overnight at 4 °C. Horseradish peroxidase (HRP)-conjugated immunoglobulin G (IgG) antibody was added and incubated for 1 h, and the specimens were analyzed through avidin and biotinylated enzyme complex (ABC) detection. Images were obtained under a ZEISS Axio Imager D2 microscope, and quantification (%Area) based on three 200× fields was performed using Nuance imaging software (version 3.0.2, PerkinElmer, Burlington, VT, USA).
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