The clinical PC and adjacent tissues were fixed with paraformaldehyde (Servicebio, Wuhan, China), embedded in paraffin (Boster, Wuhan, China), and sectioned into 4 μm thickness. After dewaxing, rehydration, and antigen retrieval with sodium citrate (Servicebio, Wuhan, China), tumor samples were blocked using H2O2 and 5% bull serum albumin (Servicebio, Wuhan, China) for 30 min at room temperature. The specimens were incubated with primary anti-TRA2A (1:100), KI67 (1:100), anti-PCNA (1:100), and anti-HIF1α (1:100) antibodies for 12 h and horseradish peroxidase-conjugated secondary antibodies (Zsjqbio.biogo; Beijing, China). Next, DAB was used to identify the antigen-antibody complex, and the samples were stained with hematoxylin to visualize the nucleus.
Paraffin
Manufactured by Boster Bio
Paraffin is a wax-like substance derived from petroleum, coal, or natural gas. It is a solid, odorless, tasteless, and inert material commonly used in various laboratory and industrial applications. Paraffin has a high melting point and is known for its ability to act as a solid support material, particularly in the field of histology and microtomy.
Automatically generated - may contain errors
2 protocols using paraffin
1
Immunohistochemical Analysis of Tumor Samples
2
Immunohistochemical Analysis of Hepatocellular Carcinoma
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The HCC and adjacent tissues were fixed, dehydrated, embedded in paraffin (Boster, Wuhan, China), and then were cut into 4-µm specimens. The specimens were stained with hematoxylin and eosin (Sigma, USA) following the manufacturer’s protocol. Specimens were deparaffinized using a graded series of xylene and rehydrated with a graded series of ethanol. Next, antigen retrieval was performed using sodium citrate and blocking was performed using H2O2 and BSA (Boster, Wuhan, China). The sections were then incubated with a primary anti-ODC1, anti-PKM2, anti-KI67, and anti-PCNA antibody for 16 h at 4°°C and then with horseradish peroxidase-conjugated secondary antibody (Boster, Wuhan, China) for 2 h at room temperature. After using a Cell and Tissue Staining Horse Radish Peroxidase (HRP) - Diaminobenzidine (DAB) Kit (Zhongshan Jinqiao Biotechnology Co. LTD; Beijing, China) to detect the presence of the secondary antibody, orthophoto microscopy was used to obtain images.
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