Specimens, including 26 CCPRCC, 30 CCRCC and 30 PRCC, were fixed in formalin and embedded in paraffin. The 4-µm thick sections were stained with the following panel of markers: CK7 (OV-TL 12/30, 1:200; Dako, Carpinteria, CA, USA); CD10 (56C6, 1:25; Novocastra, Newcastle upon Tyne, UK); AMACR (13H4, ready-to-use; Dako); CA IX (TH22, 1:100; Novocastra, Buffalo Grove, IL, USA); vimentin (Vim 3B4, 1:250); Ki67 (MIB-1, 1:200) (both from Dako, Glostrup, Denmark). Immunoreaction was performed with an automated immunostainer from Ventana (Ventana Medical Systems, Tucson, AZ, USA). The immunohistochemistry results (CK7, C10, AMACR, CA IX and RCC maker) were interpreted as negative, weak (<30% staining), moderate (30–70% staining) and strong (>70% staining). Ki67 positive cells showed stained brownish-yellow granules in the nucleus. According to the literature written by Delahunt et al (28 (link)), the area with the highest fraction of Ki67-stained cells in section was chosen at a X10 objective magnification, then it was examined at X400 objective magnification. Finally, Ki67 labeling index (Ki67 LI) was made through counting 1,000 cancer cells (percentage of nuclei showing positive staining).
Vimentin vim 3b4
Manufactured by Agilent Technologies
Sourced in United States
Vimentin (Vim 3B4) is a laboratory equipment product offered by Agilent Technologies. It is a protein marker used in cell and tissue analysis. The core function of this product is to detect and visualize the expression of the vimentin protein, which is a key component of the cytoskeleton in various cell types.
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Lab products found in correlation
Automated immunostainer, by Roche (1 mentions)
Amacr 13h4, by Agilent Technologies (1 mentions)
Cd10 56c6, by Leica (1 mentions)
Ck7 ov tl 12 30, by Agilent Technologies (1 mentions)
Ki 67 mib 1, by Agilent Technologies (1 mentions)
Cytochrome c, by Cell Signaling Technology (1 mentions)
Hsc70, by Santa Cruz Biotechnology (1 mentions)
Horseradish peroxidase conjugated secondary antibody, by Agilent Technologies (1 mentions)
Gm130 35 gm130, by BD (1 mentions)
Phospho bad, by Cell Signaling Technology (1 mentions)
Hsp90, by Cell Signaling Technology (1 mentions)
Tsg101, by Abcam (1 mentions)
α sma, by Merck Group (1 mentions)
Phospho akt, by Cell Signaling Technology (1 mentions)
β actin, by BD (1 mentions)
2 protocols using vimentin vim 3b4
1
Immunohistochemical Profiling of Renal Cell Carcinomas
2
Western Blot Analysis of Cellular and Exosomal Proteins
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Cells or exosomes were lysed in 2X Laemmli buffer (4% SDS, 20% glycerol, 10% 2-mercaptoethanol, 0.02% bromophenol blue and 120mM Tris HCl). Proteins were separated under reducing conditions in 8, 10, 12 or 15% SDS-PAGE gels, transferred onto nitrocellulose membranes, detected with AKT (pan; C67E7; 1:1000), phospho-AKT (Ser473; D9E; 1:1000), Bad (D24A9; 1:1000), phospho-Bad (Ser136/99; D25H8; 1:1000), cytochrome C (4272; 1:1000; Cell Signaling Technology, Danvers, USA), Alix (3A9; 1:500), TSG101 (4A10; 1:500; Abcam, Cambridge, UK), CD63 (Ts63; 1:500), CD81 (1.3.3.22; 1:500; Thermo Fisher, Waltham, USA), GM130 (35/GM130; 1:500; BD Biosciences, Oxford, UK), α-SMA, (1A4; 1:2000; Sigma-Aldrich, Saint Louis, USA), FN1-EDA (MAB1940; 1:2000; Merck Millipore, Burlington, USA), palladin (1E6; 1:1000; Novus Biologicals, Littleton, USA), Vimentin (Vim 3B4; 1:1000; Dako, Glostrup, Denmark) primary antibodies and horseradish peroxidase-conjugated secondary antibodies (1:4000; Dako, Glostrup, Denmark). Specific signal was visualized using the SuperSignal West Dura Chemi-luminescent detection kit (Thermo Scientific, Waltham, USA). Membranes were probed for β-actin (C4; 1:5000; BD Biosciences, San Jose, USA), HSP90 (1:1000; Cell Signaling Technology, Danvers, USA) or HSC-70, (B-6; 1:2000; Santa Cruz Biotechnology, Dallas, USA), as loading controls.
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