Cisplatin [cis-diammineplatinum(II) dichloride] was purchased from Sigma-Aldrich, LLC (St. Louis, MO). Quick coating solution used to coat the flask for human neonatal dermal fibroblast (HNDF) cells was purchased from Angio-Proteomie (Shrewsbury, MA). Monoclonal mouse anti-vimentin was purchased from Dako, Inc. (Carpinteria, CA) and rabbit anti-vimentin polyclonal antibody was purchased from Bioss, Inc. (Woburn, MA).
Monoclonal mouse anti vimentin
Manufactured by Agilent Technologies
Sourced in Denmark
Monoclonal mouse anti-vimentin is a laboratory reagent used for the detection and identification of vimentin, an intermediate filament protein. This product can be used in various immunoassays and immunocytochemical applications.
Automatically generated - may contain errors
Lab products found in correlation
Quick coating solution, by Angio-Proteomie (1 mentions)
Rabbit anti vimentin polyclonal antibody, by Bioss Antibodies (1 mentions)
Cisplatin, by Merck Group (1 mentions)
Lsab2 kit, by Agilent Technologies (1 mentions)
Envision hrp kit, by Agilent Technologies (1 mentions)
Peroxidase blocking solution, by Agilent Technologies (1 mentions)
Polyclonal rabbit anti s100, by Agilent Technologies (1 mentions)
Las af, by Leica (1 mentions)
Goat anti rabbit igg 594, by Abcam (1 mentions)
5 protocols using monoclonal mouse anti vimentin
1
Culturing Human Neonatal Dermal Fibroblasts
2
Immunohistochemical Staining of Vimentin
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Monoclonal mouse anti-Vimentin, (Code No. M 0725, Dako, Carpinteria, CA, USA), was applied as primary antibody at a 1:50 dilution on 8 μm cryosections of snap frozen spleen tissue for 30 min at room temperature. Dako Cytomation Mouse lgG'1, (Code No. X 0931, Dako), diluted to the same mouse lgG concentration as the primary mouse anti-Vimentin antibody was used as negative control and run simultaneously with sarcoid spleen sections. For visualization, DAKO LSAB2 kit (Code No. K 0679) and DAKO EnVision+/HRP kits, (Code Nos. K 4004 and K 4006) were used on the Bond autostainer for automated IHC staining.
3
Vimentin Immunohistochemistry in Paraffin Sections
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A 4-μm thick section of each chosen paraffin block sample was mounted on coated glass slides (Dako A/S, Glostrup, Denmark, K8020), dewaxed, and rehydrated. Antigen retrieval was performed with a citrate buffer (pH 6.0) in a microwave oven for 20 min. Endogenous peroxidase activity was blocked by incubating the sections in Dako REAL™ Peroxidase-Blocking solution for 5 min. Sections were then incubated with the primary antibody - Monoclonal mouse anti-vimentin (Clone V9, M0725, Dako) diluted 1:100 for 30 min. This was followed by incubation for 30 min with a ready-to-use secondary antibody (Dako REAL™ EnVision™/Horseradish Peroxidase, Rabbit/ Mouse) and with the substrate Dako REAL™ Diaminobenzidine + Chromogen for a further 10 min. Rinses were done with Dako-Cytomation Wash Buffer between each step. All of the previous steps (from antigen retrieval) were done in a Dako Autostainer. Finally, the sections were counterstained with hematoxylin, dehydrated and mounted with cover slips. A sample of connective tissue was used as a positive control, while negative controls were obtained by substitution of the primary antibody with phosphate buffered saline.
4
Immunohistochemical Profiling of Glioma Samples
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Tissue specimens or cultured cells were fixed with 4% paraformaldehyde in phosphate-buffered saline (PBS) for IHC analyses. Paraffin-embedded tissue sections from a different cohort (n = 82, WHO grade I = 58, grade II = 21, and grade III = 3) were immunostained using previously described conditions. Briefly, the sections were de-paraffinized using xylene, rehydrated and blocked with 2% goat serum. Samples were subsequently incubated with primary antibody overnight at 4°C and secondary antibody conjugated to Horseradish Peroxidase (HRP) at room temperature for 1 hour. The signals were visualized with an HRP substrate. The following primary antibodies were used: polyclonal rabbit anti-GFAP (glial fibrillary astrocytic protein) (Chemicon International, Temecula, CA), monoclonal mouse anti-EMA (epithelial membrane antigen) (Thermo Scientific, Rockford, IL), polyclonal rabbit anti-S100 and monoclonal mouse anti-vimentin (Dako, Denmark) antibodies. Transglutaminase 2 (TGM2) antibody was purchased from Abcam (Cambridge, UK). At least 4 high-power fields (HPF, 400x) were randomly photographed from the tumor region. In each HPF, individual cell counts were made by single observer and mean percentage were recorded as %/HPF.
5
Co-localization of Fibroblasts and Signaling Proteins
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To assess co-localization of fibroblasts and ROCK1 or LIMK2-activated sites in penile tissue, double immunofluorescence for ROCK1 and vimentin or phospho-LIMK2 and vimentin were performed. For immunofluorescence staining of paraffin embedded sections (2.5µm) were treated and incubated with primary antibody; monoclonal mouse anti-vimentin (1:100, Dako, Glostrup, Denmark) during two overnight periods, monoclonal rabbit anti-ROCK1 (1:10, AbCam, Cambridge, MA, USA) or rabbit polyclonal anti-LIMK2 (phospho T505) (1:100, AbCam, Cambridge, MA, USA) for two overnight periods at room temperature. After three washes in PBS, the sections were incubated with the fluorescence secondary antibody; goat antimouse IgG antibody 488 (1:400, Invitrogen, Camarillo, CA, USA) 10min for vimentin, and goat anti-rabbit IgG 594 (1:200, AbCam, Cambridge, MA, USA) 10min for ROCK1 or phospho-LIMK2. Sections were analyzed using a confocal laser fluorescencemicroscope (Leica Microsystems, Heidleberg, Germany) with single and double filter settings at 488 and 594 nm.
Images were acquired using LAS AF (Leica Microsystems, Heidleberg, Germany).
Images were acquired using LAS AF (Leica Microsystems, Heidleberg, Germany).
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