Tissue Microarray and IHC Analysis of ILF3

Surgical tissue samples was fixed with 10% formalin for 24 h at room temperature. Formalin-fixed paraffin-embedded surgical tissue samples were used for TMAs and IHC. To construct the TMAs, tissue cylinders (2 mm in diameter) were removed from each sample and selected tissue cylinders were grouped into a single array block using an Unitma Quick-Ray tissue microarrayer (cat. no. UT06; Unitma Co., Ltd). Each TMA specimen was subsequently cut into 4 µm tissue sections, which were mounted on microscope slides. IHC staining was performed as previously described (21 (link)), except an anti-ILF3 antibody (ab92355; 1:400; Abcam) was used as the primary antibody. A total of two pathologists blindly evaluated the percentage of ILF3-positive samples using the NDP.view2 software (version 2.6.13; Japan SLC, Inc.), as well as the intensity of ILF3 IHC staining. A semi-quantitative immunoreactivity scoring system was used to evaluate the staining (22 (link)). The semi-quantitative H-score (0–300) was calculated as the product of the intensity (0, negative; 1, weak; 2, moderate; 3, strong) and the percentage of ILF-3-positive samples (0–100).

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Xu Z., Huang H., Li X., Ji C., Liu Y., Liu X., Zhu J., Wang Z., Zhang H, & Shi J. (2020). High expression of interleukin-enhancer binding factor 3 predicts poor prognosis in patients with lung adenocarcinoma. Oncology Letters, 19(3), 2141-2152.

Publication 2020

ab92355

Anti antibody Antibody Formalin Microscope Paraffin Pathologists Surgical Tissue Weak

Corresponding Organization :

Other organizations : Affiliated Hospital of Nantong University, Nantong University

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Variable analysis

independent variables

Formalin fixation time (24 h)
Formalin concentration (10%)

dependent variables

Percentage of ILF3-positive samples
Intensity of ILF3 IHC staining
Semi-quantitative H-score (0-300)

control variables

Room temperature during formalin fixation
Tissue sample type (surgical tissue samples)
Tissue processing (formalin-fixed paraffin-embedded)
Tissue sectioning (4 µm)
Slide mounting
IHC staining procedure (as previously described)

positive controls

Not explicitly mentioned

negative controls

Not explicitly mentioned

Annotations

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