Experiments were performed on 4-μm paraffin-embedded tumor tissues mounted on slides. Paraffin was removed, and the sections were serially rehydrated by descending ethanol series as described above. Antigen retrieval was performed by heating the slides in EDTA buffer at 100°C for 15 min. Slides were blocked by incubating with 3% BSA (Wuhan Servicebio Technology Co., Ltd.) for 30 min at 37°C and then incubated with primary antibodies against FAM83A (1:500; BIOSS), integrin α1 (1:200; ProteinTech Group, Inc.), integrin α3 (1:1,000; ProteinTech Group, Inc.), integrin α5 (1:100; Hangzhou HuaAn Biotechnology Co., Ltd.), integrin β4 (1:100; ProteinTech Group, Inc.) and integrin β5 (1:100; BIOSS) at 4°C overnight, followed by the secondary antibodies Alexa Fluor 488-conjugated goat anti-mouse (1:400; cat. no. GB25303) or Cy-3-conjugated goat anti-rabbit (1:300; cat. no. GB21303; both from Wuhan Servicebio Technology Co., Ltd.) for 1 h at room temperature. The cell nuclei were stained with DAPI. Nikon Eclipse C1 fluorescence microscope with an integrated camera was used to visualize the fluorescence and acquire images from five representative fields of each sections (×400).
Integrin β5
Manufactured by Bioss Antibodies
Sourced in United States
Integrin β5 is a cell surface receptor that plays a role in cell-cell and cell-matrix adhesion. It is part of the integrin family of proteins, which mediate interactions between cells and the extracellular matrix. Integrin β5 forms heterodimeric complexes with various alpha integrin subunits, such as αV, and is involved in processes like cell migration, signaling, and angiogenesis.
Automatically generated - may contain errors
2 protocols using integrin β5
1
Immunofluorescence Analysis of Integrin Expression
2
Immunohistochemical Evaluation of Tumor Markers
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Paraffin‐embedded tissues were deparaffinized with dimethylbenzene followed by antigen retrieval. The tissues were blocked with normal goat serum at 37 °C for 30 min. Next, the tissues were incubated overnight at 4 °C with specific primary antibodies against integrin β5 (Bioss, Boston, MA, USA, 1 : 200), p‐FAK (CST, 1 : 600), p‐ERK (CST, 1 : 250), p‐STAT3 (CST, 1 : 200), MMP2 (Abcam, 1 : 200), HIF‐1α (Abcam, 1 : 100), and CD31 (CST, 1 : 200). Finally, the tissues were incubated with appropriate secondary antibodies and then incubated with 3, 3′‐diaminobenzidine (DAB). We evaluated the marker staining results according to a previous study [31 (link)]. The staining intensity was graded 4 stages: 0 (none), 1 (weak), 2 (moderate), and 3 (strong). The percentage of expression was graded 5 stages: 0 (< 5% staining), 1 (5–25% staining), 2 (25–50% staining), 3 (50–75% staining), and 4 (> 75% staining). The sum of both scores served as the final score. Two pathologists performed the scoring analyses according to the above criteria.
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