The streptavidin–biotin peroxidase complex method was utilized for immunohistochemical studies with antibodies to epithelial membrane antigen (EMA; Clone E29, Ventana, monoclonal), somatostatin receptor 2 (SSTR2; Clone UMB1, Abcam, 1:500, monoclonal), smooth muscle actin (SMA; Clone 1A4, Cell Marque, 1:200 monoclonal), Ki-67 (MIB-1; Clone MIB-1, Dako,1:200 monoclonal), CD34 (Clone QBEnd-10, Ventana, monoclonal), ERG (Clone EPR3864, Ventana; monoclonal), CD68 (Clone KP1, Ventana, monoclonal). Antibodies were obtained and applied at ready-to-use dilutions unless stated otherwise.
Clone kp1
Manufactured by Roche
Sourced in United Kingdom
The Clone KP1 is a laboratory equipment product developed by Roche. It is a device designed for cloning and amplification of DNA sequences. The core function of the Clone KP1 is to facilitate the replication of genetic material for various scientific and research applications.
Automatically generated - may contain errors
Lab products found in correlation
Clone mib 1, by Agilent Technologies (1 mentions)
Clone umb1, by Abcam (1 mentions)
Clone qbend 10, by Roche (1 mentions)
Clone epr3864, by Roche (1 mentions)
Clone pg m1, by Agilent Technologies (1 mentions)
Cd163 clone mrq 26, by Roche (1 mentions)
Cd4 for t helper, by Roche (1 mentions)
Clone sp57, by Roche (1 mentions)
3 protocols using clone kp1
1
Immunohistochemical Staining of Tumor Markers
2
Immunohistochemical and In-Situ Hybridization Analysis
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For immunohistochemistry, 4μ-thick sections obtained from formalin-fixed/paraffin-embedded tissues were placed on positive-charged electrostatic slides (Isotherm Technical Laboratory Glass Materials). All sections were then placed on a fully automated immunohistochemical staining machine (Ventana, Benchmark, XT IHC/ISH). UltraView Universal DAB Detection Kit compatible with the device was used for IHC staining. Monoclonal mouse primary antibodies CD163 (clone MRQ-26, prediluted, VENTANA, Cat. No: 760-4437) and CD68 (clone KP-1, prediluted, VENTANA, Cat. No: 790-2931and clone PG-M1, prediluted, DAKO, Code: IS613, prediluted, Cat. No: 760-4437) were applied and staining was completed according to the standard procedures. To detect EBV-RNA with an EBER (Epstein-Barr virus-encoded RNA) probe via the CISH method, 4μ-thick tissue sections placed on positive-charged electrostatic slides were also used. All tests were performed with a fully automated ISH machine (Ventana, Benchmark XT, IHC/ISH), by using EBER 1 DNP Probe (Regulatory status: ASR; Cat. No: 760-1209), compatible with the machine, and the Ultraview AP Red ISH Kit for EBV RNA signaling, and staining was completed according to the standard procedures.
3
Immunohistochemical Profiling of Tonsil Cells
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IHC was performed on 4 µm serial FFPE tonsil sections to characterize cell populations with the following antibodies: CD8 for cytotoxic T lymphocyte (CTL) (clone SP57, Ventana Roche, Tucson, USA), Granzyme B (GrB) for activated cytotoxic cells (clone GB7, AbD Serotec, Oxford, UK), CD68 for macrophages (clone KP-1, Ventana Roche), IL10 for anti-inflammatory cytokine-productive cells (Abcam), Foxp3 for regulatory T lymphocytes (Treg) (Abcam, Cambridge, UK), PD1 (CD279) for T cell-negative regulator (AbD Serotec), CD56 for NK cells (Leica, Buffalo, IL, USA) and CD4 for T helper (Th) (Ventana Roche). Lymph node reactive hyperplasia tissue was used as positive control. Negative controls for each case consisted in substituting the primary antibody with antibody dilution buffer and an isotype control. The stains were developed using diaminobenzidine (DAB).
Given the fact that we previously characterized viral antigen expression in lymphocytes at germinal center (GC), interfollicular (IF) and subepithelial (SubEp) regions, the same approach was used for microenvironment characterization at those three histological regions [19, (link)20] (link).
All cell markers were observed and counted by two pathologists in serial slides on the basis of the best-preserved areas around LMP1+ and LMP1-zones. The results were expressed as immunopositive cells nº/100 total cell nº.
Given the fact that we previously characterized viral antigen expression in lymphocytes at germinal center (GC), interfollicular (IF) and subepithelial (SubEp) regions, the same approach was used for microenvironment characterization at those three histological regions [19, (link)20] (link).
All cell markers were observed and counted by two pathologists in serial slides on the basis of the best-preserved areas around LMP1+ and LMP1-zones. The results were expressed as immunopositive cells nº/100 total cell nº.
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