Immunohistochemical Analysis of Cell Markers

The tissue sections were dewaxed and debenzylated according to previous study [37 (link)]. and then incubated with rabbit anti-Ki67 (Abcam, USA), rabbit anti-HIF-1α (Abcam, USA), rabbit anti-NCR1 (Abcam, USA) and rabbit anti-CD3 (CST, USA) antibodies overnight at 4 °C. After washing, the bound antibodies were detected using horseradish peroxidase-conjugated secondary antibody (DAKO, Denmark) and diaminobenzidine (DAKO, Denmark), followed by counterstaining with hematoxylin (Keygen Biotech, Nanjing, China). The images were viewed under a fluorescence microscope (LEICA DMI 4000B, Solms, Germany) and were analyzed by Leica application suite software (version 4.0).

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Jia C., Zhang F., Lin J., Feng L., Wang T., Feng Y., Yuan F., Mai Y., Zeng X, & Zhang Q. (2022). Black phosphorus-Au-thiosugar nanosheets mediated photothermal induced anti-tumor effect enhancement by promoting infiltration of NK cells in hepatocellular carcinoma. Journal of Nanobiotechnology, 20, 90.

Publication 2022

rabbit anti-Ki67 rabbit anti-HIF-1α rabbit anti-NCR1 horseradish peroxidase-conjugated secondary antibody hematoxylin DMI 4000B

Anti cd3 Antibodies Antibody Fluorescence microscope Hematoxylin Horseradish peroxidase Rabbit Tissue

Corresponding Organization :

Other organizations : Third Affiliated Hospital of Sun Yat-sen University, Sun Yat-sen University

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Variable analysis

independent variables

Rabbit anti-Ki67 antibody
Rabbit anti-HIF-1α antibody
Rabbit anti-NCR1 antibody
Rabbit anti-CD3 antibody

dependent variables

Protein expression of Ki67, HIF-1α, NCR1, and CD3 in tissue sections

control variables

Tissue sections were dewaxed and debenzylated according to previous study [37]
Tissue sections were incubated with antibodies overnight at 4 °C
Bound antibodies were detected using horseradish peroxidase-conjugated secondary antibody and diaminobenzidine
Tissue sections were counterstained with hematoxylin
Images were viewed under a fluorescence microscope and analyzed by Leica application suite software

controls

Positive control: Not explicitly mentioned.
Negative control: Not explicitly mentioned.

Annotations

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