After the treatments, mucosal biopsies were fixed in buffered formalin, embedded in paraffin and cut into 5 μm‐thick serial sections. According to the manufacturer's instructions, after heat‐mediated antigen retrieval, the tissue was formaldehyde fixed and blocked with serum. The tissue was incubated with the primary antibodies anti‐S100B (1:50 v/v) or anti‐wtp53 (1:50 v/v), (both from Abcam) for 20 minutes. In supplementary experiments, slices were incubated with anti‐MAC387 (1:100 v/v) (Abcam). After three 5‐minute washes, the secondary antibody was added and the samples were incubated at room temperature for 20 minutes. The streptavidin‐HRP detection system (Chemicon Int.) was added and samples were incubated at room temperature. After three 5‐minute washes, 50 μL of chromogen was added and the reaction terminated after 1 minute in water. Sections were then counterstained with haematoxylin‐eosin at room temperature. Negative controls were performed by omitting the primary antibody. Slides were thus analysed with a microscope (Nikon Eclipse 80i by Nikon Instruments Europe), and images were captured at 20× magnification by a high‐resolution digital camera (Nikon Digital Sight DS‐U1). Results were expressed as positive cells per μm2.
Anti s100b
Manufactured by Abcam
Sourced in United Kingdom, United States
Anti‐S100B is a laboratory reagent used to detect and quantify the S100B protein. S100B is a calcium-binding protein involved in various cellular processes. This antibody product can be used in techniques such as Western blotting, immunohistochemistry, and ELISA to measure S100B levels in biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Anti s100, by Abcam (2 mentions)
Anti map2 antibody, by Cell Signaling Technology (2 mentions)
Anti aldh1l1, by Merck Group (2 mentions)
Anti cd11b, by Thermo Fisher Scientific (2 mentions)
Anti cd68, by Cell Signaling Technology (2 mentions)
Paraformaldehyde aqueous solution, by Electron Microscopy Sciences (2 mentions)
Anti p tau antibody, by Thermo Fisher Scientific (2 mentions)
Anti s100a6, by Cell Signaling Technology (2 mentions)
Digital sight ds u1, by Nikon (1 mentions)
Anti mac387, by Abcam (1 mentions)
Eclipse 80i, by Nikon (1 mentions)
Anti nf200, by Abcam (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Wuhan Servicebio Technology (1 mentions)
Fluorescence microscope, by Leica (1 mentions)
Bovine serum albumin (bsa), by Merck Group (1 mentions)
Goat serum, by Merck Group (1 mentions)
Gabapentin neurontin, by Pfizer (1 mentions)
Anti ng2, by Santa Cruz Biotechnology (1 mentions)
Sybr select master mix, by Thermo Fisher Scientific (1 mentions)
High capacity cdna reverse transcription kit, by Thermo Fisher Scientific (1 mentions)
7 protocols using anti s100b
1
Immunohistochemical Staining of Mucosal Biopsies
2
Immunofluorescence Staining of Paraffin Sections
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Paraffin sections, prepared as described above, were deparaffinized and incubated with anti‐NF200 (1:100, Abcam, UK) antibody and anti‐S100B (1:100, Abcam, UK) antibody, respectively. The sections were incubated overnight, and rinsed, and cell nuclei were labeled using DAPI (Servicebio, Wuhan, China). Observation and image acquisition of the sections were conducted using a Leica fluorescence microscope.
3
Immunofluorescent Staining of Cells and 3D Cultures
4
Immunofluorescent Visualization of Tau and S100B
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For immunofluorescence, U-251 MG or differentiated SH-SY5Y cells were first fixed with 4% paraformaldehyde (Cat. AP.EM-15710, Electron Microscopy Sciences, PA, USA), permeabilized with 0.1% Triton X-100 (Cat. X100RS, Sigma-Aldrich, St. Louis, MO, USA) and blocked with blocking buffer (2% bovine serum albumin) (Cat. A7030, Sigma-Aldrich, St. Louis, MO, USA) and 5% goat serum (Cat. X0907, Agilent Technologies, Santa Clara, CA, USA). Anti-tau (Cat. sc-58860, Santa Cruz Biotechnology, Dallas, TX, USA) and anti-S100B (Cat. ab-52642, Abcam, Cambridge, UK) antibodies were added at a dilution of 1/100 on blocking buffer and incubated overnight at 4 °C. After washing with phosphate buffer saline (PBS), anti-mouse IgG Alexa Fluor 647-conjugated (Cat. A-21235, Thermo Fisher Scientific, Waltham, MA, USA) and anti-rabbit IgG Alexa Fluor 488-conjugated (Cat. O-6381, Thermo Fisher Scientific, Waltham, MA, USA) secondary antibodies were added at a 1/500 dilution in blocking buffer and incubated for 1 h in a dark humid chamber. For nuclei visualization, cells were counterstained with DAPI (Cat. D9542, Sigma-Aldrich, St. Louis, MO, USA) at 0.5 µM for 10 min. Coverslips were then mounted in slides with mounting medium. Coverslips were sealed and stored at 4 °C until acquisition of fluorescence microscopy images.
5
Immunohistochemical Analysis of CNS Markers
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Antibodies were purchased from DAKO (policlonal anti-Glial Fibrillary Acidic Protein [GFAP]), Sigma (monoclonal anti-GFAP, anti-S100B), Abcam (anti-Iba-1), Millipore-Chemicon (anti-NG2), and Santa Cruz (anti-thrombospondin-1 [TSP-1]). Secondary fluorescent or biotinilated antibodies were from Jackson Immunoresearch and Sigma (St. Louis, MO, USA) respectively. Gabapentin (Neurontin) was from Pfizer (Buenos Aires, Argentina). For Quantitative RT-PCR, the Quick-RNA™ MiniPrep (ZYMO Research,#R1054, Irvine, CA, USA.) was used for RNA extraction, the High-Capacity cDNA Reverse Transcription Kit (Applied Biosystems, Foster City, CA, USA) was used for retrotranscription, and the SYBR® Select Master Mix (Life Technologies, Carlsbad, CA, USA) for Real Time PCR.
6
Immunohistochemical Analysis of Mouse Brain
7
Immunofluorescent Staining of Cells and 3D Cultures
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For immunofluorescent stains, we rinsed the cells and 3D cultures twice with PBS (phosphate buffered saline). Cells were then fixed through a RT 15–30 min incubation in fresh 4% paraformaldehyde aqueous solution (157–4, ElectronMicroscopy Sciences) followed by rinsing twice with PBS. Cells were permeabilized through incubation in 0.1% Triton X-100 in PBST (phosphate buffered saline with 0.1% tween®20) for 15 minutes at RT. Cell on-specific binding was blocked through overnight incubation in 3% human serum albumin in PBST at 4 °C. After a 24-hours incubation with the primary antibody solutions at 4°C, the cells were washed five times. The following antibodies (and dilutions) were used: anti-p-tau antibody (1:40, AT8, Thermo Scientific, MN1020); anti-PHF (1:1000, A gift from P. Davies, Albert Einstein College of Medicine); anti-GFAP antibody (1:500, Neuromap, N206A/8), anti-MAP2 antibody (1:200, Cell Signaling Technology, 4542); anti-CD68 (1:100, Cell Signaling, 76437); anti-cd11b (1:100, Life Technologies, NB110–89474); anti-S100 (1:400, Abcam, ab868); anti-S100A6 (1:200, Cell Signaling, D3H3W); anti-S100B (1:500, Abcam, Ab218515); anti-ALDH1L1 (1:200, EMD Millipore, MABN495).
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