Immunohistochemistry (IHC) assay was designed to analyze the expression of nuclear proliferation marker (Ki‐67), Cleaved‐caspase 3, E‐cadherin and N‐cadherin in the transplanted tumors from xenograft tumor model assay as instructed.26 The tumors were cut into the 4‐µm‐thick sections, and embedded into paraffin, followed by the heating at 60°C. Twenty minutes later, the sections were deparaffinized and hydrated with xylene (Millipore) as well as ethanol (Millipore). After that, the sections were dipped in the sodium citrate (Millipore) and heated to conduct the antigen retrieval. After being immersed in Hydrogen Peroxide (Millipore) for 10 min, the sections were incubated with the primary antibodies against Ki‐67 (AF0198; 1:100; Affinity), Cleaved‐caspase 3 (AF7022; 1:100; Affinity), E‐cadherin (AF0131; 1:100; Affinity) and N‐cadherin (AF4039; 1:100; Affinity) as well as secondary antibodies (S0001; 1:200; Affinity), respectively. Hematoxylin (Millipore) was incubated with the tissues, and protein expression was analyzed under a CX31‐LV320 microscope (Olympus).
Cx31 lv320 microscope
Manufactured by Olympus
Sourced in Japan
The CX31-LV320 is a compact and versatile microscope designed for routine laboratory use. It features a monocular observation tube and supports multiple magnification levels to accommodate a variety of sample types. The microscope is equipped with an LED illumination system and a range of objectives to provide clear, high-quality images.
Automatically generated - may contain errors
Lab products found in correlation
Ihc assay kit, by Phygene (2 mentions)
Xylene, by Merck Group (1 mentions)
Sodium citrate, by Merck Group (1 mentions)
Hydrogen peroxide, by Merck Group (1 mentions)
Hematoxylin, by Merck Group (1 mentions)
Af0131, by Affinity Biosciences (1 mentions)
Ethanol, by Merck Group (1 mentions)
Af4039, by Affinity Biosciences (1 mentions)
Af0198, by Affinity Biosciences (1 mentions)
S0001, by Affinity Biosciences (1 mentions)
Af7022, by Affinity Biosciences (1 mentions)
Ab16667, by Abcam (1 mentions)
3 protocols using cx31 lv320 microscope
1
Immunohistochemical Analysis of Xenograft Tumors
2
Immunohistochemistry of A549 Tumor Samples
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IHC assay was performed on the primary tumors from A549 cells according to the standard method [26 (link)]. In brief, 4-µm-thick sections embedded into paraffin were heated and deparaffinized with xylene. The slides were incubated with H2O2 and primary antibodies specific to HIF1A (1:200; Cusabio Biotech, Wuhan, China), proliferating cell nuclear antigen (Ki67; 1:200; Cusabio Biotech) and Cleaved Caspase-3 (1:200; Cusabio Biotech). Subsequent steps were carried out using IHC assay kit (Phygene, Fuzhou, China) as instructed. CX31-LV320 microscope (Olympus) was utilized to capture images. The relative expression of the three proteins was calculated based on the percentage of stained cells and intensity of immunostaining as instructed [27 (link)].
3
Evaluation of KDM1A and Ki-67 Expression
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The expression of KDM1A and nuclear proliferation marker (Ki-67) was evaluated in this part following the published methods [18 ]. Briefly, partial xenograft was sectioned to 4 μm thickness, and embedded into paraffin, followed by deparaffinization and hydration. Afterward, these slides were incubated with the primary antibody specific to KDM1A (#PA5-17361; 1:200; Thermo Fisher Scientific) or Ki-67 (#ab16667; 1:200; Abcam). Subsequent steps were conducted with an IHC assay kit (Phygene) following the guidebook. CX31-LV320 microscope (Olympus, Tokyo, Japan) was used to capture the staining results.
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