Standard procedures were followed to conduct an IHC assay of TBCE (Abcam, ab224673, Cambridge, UK), γH2AX (Servicebio, GB111841, Guangzhou, China), Ki67 (Servicebio, GB111499, Guangzhou, China), and cleaved-Caspase-3 (Servicebio, GB11532, Guangzhou, China) in the tumor sections. Briefly, the sections were heated to 60 °C for 2 h, deparaffinized with xylene, and then the xylene was removed using alcohol. The antigenic epitopes of tissue sections were immediately retrieved with general antigen repair solution (Beyotime, P0088, Shanghai, China) and blocked with antigen-blocking horse serum (HyClone, SH30074, Los Angeles, America). Then, the tissue sections were co-incubated with the primary antibody in a refrigerator at 4 °C for 12 h. After rinsing the remaining primary antibody on each section, the corresponding antibody was incubated with HCC tissue slices, and nuclei were counter stained with hematoxylin. The IHC score was calculated based on the staining intensity and stained ratio20 (link). For specific information about antibodies, see Supporting Information Table S1 .
γh2ax
Manufactured by Wuhan Servicebio Technology
Sourced in United Kingdom, China
The γH2AX is a lab equipment product that detects and quantifies DNA double-strand breaks. It functions by identifying the phosphorylation of the H2AX histone variant, which is a marker of DNA damage response.
Automatically generated - may contain errors
Lab products found in correlation
Gb11532, by Wuhan Servicebio Technology (1 mentions)
P0088, by Beyotime (1 mentions)
Gb111841, by Wuhan Servicebio Technology (1 mentions)
Gb111499, by Wuhan Servicebio Technology (1 mentions)
Panoramic slice scanner, by 3DHISTECH (1 mentions)
Pchk1, by Abcam (1 mentions)
Senp5, by Proteintech (1 mentions)
Rad51, by Abcam (1 mentions)
3 protocols using γh2ax
1
Immunohistochemical Profiling of Tumor Markers
2
Immunohistochemical Analysis of DNA Damage Markers
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Immunohistochemistry was performed as previously described (Yi et al. 2020 (link)). Poly (ADP-ribosylation) (PAR, Novus Biologicals, US. Product number: NBP2-89039) and gamma H2A.X (p S139) (γH2AX, Servicebio, Wuhan, China, Product number: GB111841) were used as the primary antibody in the experimental protocol. Nuclei of cells were stained blue with hematoxylin. Positive staining of diaminobenzidine (DAB) was brownish yellow.
3
Immunohistochemical Analysis of DNA Damage Markers
Check if the same lab product or an alternative is used in the 5 most similar protocols
Immunohistochemistry was performed as described. Briefly, 5-micron thick formalin-fixed paraffin embedded human tissue sections were stained with the SENP5 (#19529-1-AP, Proteintech, USA), Ki67 (GB111499, Servicebio, China), γ-H2AX(GB111841, Servicebio, China), RAD51 (#ab1837, abcam, US) or p-CHK1 (#ab47318, abcam, US) antibody per manufacturer’s instructions. TUNEL staining was conducted to evaluate apoptosis in resected tissue according to the manufacturer’s instructions (Servicebio, Wuhan, China). Stained slides were digitized using the panoramic slice scanner (3DHISTECH, Hungary) with a 40× objective. Three fields of view per section were used to determine the mean and standard error of the mean of positively staining cells.
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Immunohistochemistry was performed as described. Briefly, 5-micron thick formalin-fixed paraffin embedded human tissue sections were stained with the SENP5 (#19529-1-AP, Proteintech, USA), Ki67 (GB111499, Servicebio, China), γ-H2AX(GB111841, Servicebio, China), RAD51 (#ab1837, abcam, US) or p-CHK1 (#ab47318, abcam, US) antibody per manufacturer’s instructions. TUNEL staining was conducted to evaluate apoptosis in resected tissue according to the manufacturer’s instructions (Servicebio, Wuhan, China). Stained slides were digitized using the panoramic slice scanner (3DHISTECH, Hungary) with a 40× objective. Three fields of view per section were used to determine the mean and standard error of the mean of positively staining cells.
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