Immunohistochemical (IHC) staining for S-100 protein (Dako, Santa Clara, CA, USA), cytokeratin (Dako) and CD68 (Dako) was performed on formalin-fixed, paraffin-embedded tissue for all cases to validate the diagnosis of hibernoma. All IHC analyses were performed using the Ventana Benchmark XT automated staining system (Ventana Medical Systems, Tucson, AZ, USA).
S 100 protein
Manufactured by Agilent Technologies
Sourced in Denmark, United States, Germany
The S-100 protein is a calcium-binding protein that is commonly used as a biomarker in various laboratory applications. It is a small, acidic protein that is primarily found in the cytoplasm of glial cells and certain neuronal cells in the central nervous system. The S-100 protein plays a role in the regulation of a variety of intracellular and extracellular activities, including cell growth, differentiation, and motility.
Automatically generated - may contain errors
Lab products found in correlation
Vimentin, by Agilent Technologies (3 mentions)
Desmin, by Agilent Technologies (3 mentions)
Hmb45, by Agilent Technologies (2 mentions)
Benchmark xt system, by Roche (2 mentions)
Cd31 clone jc70a, by Agilent Technologies (2 mentions)
Desmin clone d33, by Agilent Technologies (2 mentions)
Cd34 clone bi 3c5, by Zytomed Systems (2 mentions)
Ttf1 clone 8g7g3 1, by Zytomed Systems (2 mentions)
Ki 67 clone mib 1, by Agilent Technologies (2 mentions)
Glial fibrillary acidic protein (gfap), by Agilent Technologies (2 mentions)
Ventana benchmark xt, by Roche (1 mentions)
Bond max, by Leica (1 mentions)
Synaptophysin, by Agilent Technologies (1 mentions)
Chromogranin a, by Agilent Technologies (1 mentions)
α smooth muscle actin αsma, by Thermo Fisher Scientific (1 mentions)
Melan a, by Agilent Technologies (1 mentions)
Cathepsin k, by Abcam (1 mentions)
Stat6, by Santa Cruz Biotechnology (1 mentions)
Alpha smooth muscle actin clone 1a4, by Agilent Technologies (1 mentions)
Erg epr3864, by Roche (1 mentions)
17 protocols using s 100 protein
1
Immunohistochemical Validation of Hibernoma
2
Analyzing Oral Squamous Cell Carcinoma
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Formalin-fixed, paraffin-embedded tissue blocks were provided by our Department of Pathology. Three pathologists examined all resected specimens to confirm the histopathological features. The stage grouping of the tumors was classified according to the TNM classification of malignant tumors, seventh edition [25 ]. The mode of invasion was examined at the host/tumor interface as previously defined by Yamamoto et al. [26 (link)]. Each patient was assigned an Eastern Cooperative Oncology Group performance status (ECOG PS) score at the time of diagnosis [27 (link)]. Non-carcinomatous tissue and recurrent OSCC tissue were obtained from non-tumor portions of the surgically resected specimens and metastatic regional lymph nodes or locally recurrent oral mucosa corresponding to each primary site, respectively. These tissues were stained with hematoxylin and eosin (H&E), Elastica van Gieson (EVG), or immunohistochemistry preparations of sequential sections. EVG, immunohistochemical Podoplanin (D2-40; Nichirei Bioscience Co., Tokyo, Japan; diluted 1:1), and S-100 protein (Dako, Glostrup, Denmark; diluted 1:900) staining very clearly revealed vascular invasion (v), lymphatic vessel invasion (ly), and perineural involvement (ne), respectively, as previously described [28 (link)].
3
Immunohistochemical Staining Optimization
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Immunostaining was performed using a BOND‐MAX autoimmunostainer (Leica Microsystems, Wetzlar, Germany). Deparaffinized and rehydrated sections were subjected to endogenous peroxidase blocking. After heating in antigen unmasking solution, slides were incubated with one the following antibodies: S‐100 protein (Dako, Santa Clara, CA, USA), mammaglobin (304‐1A5, 1:100, Dako), DOG1 (SP31, 1:50, Thermo Scientific, Cheshire, UK) and α‐amylase (1:50, Nordic Immunology, Tiburg, the Netherlands). Color development was carried out using 3,3′‐diaminobenzidine tetrahydrochloride, and slides were counterstained with hematoxylin. Appropriate tissue sections were used as positive controls, and procedures without primary antibodies were used as negative controls.
4
Comprehensive Immunohistochemical Profiling
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HE staining: All specimens were fixed by 10% neutral formalin, regularly dehydrated, paraffin-embedded, sectioned into 3-μm sections, and processed for HE staining. IHC staining: IHC staining was performed using the EnVision two-step strategy. Primary antibody information: CD117, HMB45, Desmin, synaptophysin, chromograninA, S100 protein, wide-spectrum cytokeratin, Ki67, and vimentin (DAKO); Bcl2, P53, CD56, CD57 (LEICA); muscle specific actin (MSA), CD34, H-caldesmon (LongIsland); SOX10, INI1, DOG-1 (Gene Tech); somatostatin receptor 2 (SSTR2), somatostatin receptor 5 (SSTR5) (Abcam); α-smooth muscle actin (α-SMA) (Thermo); ATRX (Sigma); calponin, Collagen type IV (MXB Biotechnologies); BRAF V600E (ZSGB Biotechnology). The dilutions, clone and sources of these antibodies were listed in Table 1 .
5
Comprehensive Immunohistochemistry Panel for Tumor Diagnosis
6
Immunohistochemical Analysis of Biomarkers
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Immunohistochemical analysis was performed using the EnVision Plus detection system (DAKO, Carpinteria, CA, USA) with positive and negative controls. Antibodies for the experiments included CD34 (EP88, ready-to-use; ZSGB-Bio), SMA (UMAB237, ready-to-use; ZSGB-Bio), Ki-67 (clone MIB-1, 1:100; Dako), desmin (D33, 1:100; Dako), S-100 protein (4C49, 1:100; Dako), myogenin (F5D, 1:50; ZSGB-Bio), Bcl-2 (EP36, 1:200; ZSGB-Bio), CD99 (EP8, ready-to-use; ZSGB-Bio), p16 (16P04/IC2, ready-to-use; Dako), p63 (4A4, 1:400; Dako), cytokeratin (AE1/AE3, 1:100; Dako), and EMA (E29, 1:100; Dako).
7
Immunohistochemical Analysis of Tissue Samples
8
Immunohistochemical Staining of Melanocytic Markers
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Available tumor tissues and derived cell lines were stained with antibodies against Melan A, S-100 protein, tyrosinase, HMB 45 (all antibodies from Dako, Carpinteria, CA). The Envision Plus detection system (Dako) for all antibodies was used. Appropriate positive and negative controls were included.
9
Characterization of Undifferentiated Adipose-Derived Stem Cells
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Immunostaining was performed on undifferentiated stem cells (uADSCs) at passage 2 cultured on LabTek™ (Nunc) slides. After blocking with normal serum, the primary antibodies were applied for 2 h at room temperature CD73 (1:100; BD Biosciences), CD90 (1:1000; Millipore), CD105 (1:20; Abcam Ltd) and CD34 (1:50; Santa Cruz Biotechnology). After rinsing in phosphate-buffered saline, either secondary goat anti-mouse or donkey anti-goat Alexa Fluor 488 conjugated antibodies (1:300; ThermoFisher) were applied for 1 h at room temperature in the dark. The slides were then cover-slipped with ProLong mounting media containing 4–6-diamido-2-phenylindole (DAPI; ThermoFisher). The specificity of staining was tested by omission of the primary antibodies. To confirm multi-potency the uADSCs were treated with either adipogenic or osteogenic supplements according to the protocol described by the manufacturer (Rat Mesenchymal Stem Cell Functional Identification Kit, R & D Systems). Stem cells which were induced to a Schwann cell-like phenotype were immunostained with Sox-10 (1:200; R&D Systems), S100 protein (1:2000; Dako) and glial fibrillary acidic protein (GFAP 1:1000; Dako) antibodies. For comparison, uADSCs and primary Schwann cells were stained under identical conditions.
10
Immunohistochemical Characterization of Canine Melanocytes
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Collected tumor tissues from dogs and xenograft were fixed in 10% buffered formalin solution (Sigma-Aldrich, St Louis, MI) and paraffin embedded for routine pathological analysis. Immunohistochemical staining was carried out on 4- to 6-μm-thick slides using an automated protocol developed for the Discovery XT automated slide staining system (Ventana Medical Systems, Inc.). Alongside haematoxylin–eosin staining (HE), the following commercially available antibodies were used for the phenotypical characterization of melanocytes: Melan-A (Thermo Fisher Scientific, Waltham, MA), S-100 protein (Dako, Agilent Technologies, Denmark), Vimentin, and cytokeratin (Biogenex Laboratories, San Ramon, CA) (see Additional file 1 ). Tumor sections were deparaffinized and incubated for 1 h with the appropriate antibody before incubation with Discovery UltraMap anti-Rabbit (760–4315, Roche, Basel, Switzerland) or anti-mouse horseradish peroxidase (HRP) (760–4313, Roche, Basel, Switzerland) secondary antibodies and the Discovery ChromoMap DAB kit reagents (760–159, Roche, Basel, Switzerland) (Additional file 1 ).
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