Protocol detail

Immunohistochemical Analysis of PLIN2 and Immune Markers

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The protocol of IHC of formalin-fixed paraffin-embedded sections was performed as previously described (19 (link)). Anti-PLIN2 (ab181452, Abcam, Waltham, MA, USA) was used at a dilution ratio of 1:300, and the serial sections were incubated with primary antibodies such as anti-CD4 (ZSGB-BIO, ZM-0418), anti-CD8 (ZSGB-BIO, ZA-0508), anti-CD19 (ZSGB-BIO, ZM-0038), anti-CD56 (ZSGB-BIO, ZM-0057), anti-CD68 (ZSGB-BIO, ZM-0464), and anti-FOXP3 (ab253297, Abcam). We used PBS to replace the primary antibody as negative control. The IHC staining results of PLIN2 was assessed by ImageJ software (20 (link), 21 (link)). Digital photos were acquired from the tumor center and invasive front, and the average values were calculated for further analysis (22 (link)).

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He Y., Dong Y., Zhang X., Ding Z., Song Y., Huang X., Chen S., Wang Z., Ni Y, & Ding L. (2022). Lipid Droplet-Related PLIN2 in CD68+ Tumor-Associated Macrophage of Oral Squamous Cell Carcinoma: Implications for Cancer Prognosis and Immunotherapy. Frontiers in Oncology, 12, 824235.

Publication 2022

ab181452 ab253297

Antibodies Antibody Digital Dilution Embedded paraffin Formalin Tumor

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Variable analysis

independent variables

Anti-PLIN2 antibody dilution ratio (1:300)

dependent variables

PLIN2 immunohistochemistry (IHC) staining results assessed by ImageJ software
Digital photos of tumor center and invasive front

control variables

Formalin-fixed paraffin-embedded sections
Primary antibodies: anti-CD4, anti-CD8, anti-CD19, anti-CD56, anti-CD68, and anti-FOXP3
Negative control: PBS replacing primary antibody

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