Paraffin-embedded SACC tissue sections were immersed in xylene for 15–20 min to remove the paraffin then in decreasing concentration of ethanol to be rehydrated. 3 % H2O2 was used to block the endogenous peroxidase. Next, tissue sections were put into citrate buffer solution (PH 6.0) in a 700 W microwave for 15 min and cooled naturally to room temperature. After 20 min blocking by 5 % normal goat serum, the rabbit anti-DEC2 (1:150; Proteintech) was added to the sections to incubate overnight at 4 °C. The next day, rewarmed the slides naturally to room temperature. Sections were washed with PBS, and incubated first with biotinylated anti-mouse/rabbit IgG for 1 h and then with streptavidin-biotin peroxidase for 30 min. DAB was added to detect the primary antibody and hematoxylin was used to stain the nucleus. For the ICC staining, cells were fixed by 4 % PFA for 15 min. after washing 3 times with PBS, the cells were incubated with Triton X-100 for 20 min. The rest of the process was similar to the IHC staining.
Rabbit anti dec2
Manufactured by Proteintech
Rabbit anti-DEC2 is a primary antibody that specifically binds to the DEC2 protein. DEC2 is a transcription factor involved in the regulation of circadian rhythms. This antibody can be used to detect and study the DEC2 protein in various research applications.
Automatically generated - may contain errors
2 protocols using rabbit anti dec2
1
Immunohistochemical and Immunocytochemical Analysis
2
Immunohistochemical Analysis of DEC2, Ki-67, and NR2F1
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Paraffin-embedded sections were cut into 4um and deparaffinized in xylene and rehydrated, and endogenous peroxidase was blocked with 3%H2O2. Antigen retrieval was accomplished by 0.01 mol/L citrate buffer solution (pH 6.0) in a 700 W microwave oven for 15 min. After incubation with 5% normal goat serum for 20 min, the slides were exposed overnight at 4 °C to the rabbit anti-DEC2 (1:150; Proteintech), rabbit anti-Ki-67(1:800; Proteintech), rabbit anti-NR2F1 (1:200; Proteintech). Sections were then incubated with biotinylated goat anti-rabbit IgG (Zhongshan Goldenbridge Biotechnology) for 1 h, and streptavidin-peroxidase for 30 min. The 0.02% diaminobenzidine tetrahydrochloride was used as a chromogen, and the slides were counterstained with hematoxylin. The percentage of positive cells was estimated using an image analysis system (Leica).
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