For indirect immunofluorescence analysis, cells were fixed in 4% paraformaldehyde, permeabilized with 0.2% Triton-X100 and incubated with the following primary antibodies: mouse monoclonal α-YAP (SC-101199, Santa Cruz Biotechnology, inc.; 1:50); mouse monoclonal α-Snail (L70G2, Cell Signaling Technology; 1:50); rabbit monoclonal α-Vimentin (ab92547, Abcam, Cambridge, UK; 1:400); mouse monoclonal α-E-cadherin (BD 610182, BD Biosciences Pharmingen, Palo Alto, CA, USA; 1:50); mouse monoclonal α-STAT3 (124H6, Cell Signaling; 1:50). Secondary antibodies: α-mouse Alexa-Fluor 594 and α-rabbit Alexa-Fluor 488 (Molecular Probes, Eugene, OR, USA; 1:400). Nuclei were stained with DAPI (Calbiochem Merck, Darmstadt, Germany). Preparations were examined under Nikon Eclipse fluorescent microscope equipped with a CCD camera (Nikon Inc.). Digital images were acquired by Nikon NIS elements software (Nikon Corporation) and processed with Adobe Photoshop 7 software (Adobe Systems, Mountain View, CA). The same enhanced color levels were applied for all channels.
α vimentin
Manufactured by Abcam
Sourced in United Kingdom
α-Vimentin is a protein that plays a role in the structural integrity and organization of the cytoskeleton. It is a member of the vimentin family of intermediate filament proteins.
Automatically generated - may contain errors
Lab products found in correlation
Alexa fluor 488 α rabbit, by Thermo Fisher Scientific (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions)
Ccd camera, by Nikon (1 mentions)
α e cadherin, by BD (1 mentions)
Nis elements software, by Nikon (1 mentions)
α stat3, by Cell Signaling Technology (1 mentions)
Supersignal west pico, by Thermo Fisher Scientific (1 mentions)
α e cadherin, by Abcam (1 mentions)
α gapdh, by Abcam (1 mentions)
2 protocols using α vimentin
1
Indirect Immunofluorescence Analysis of EMT Markers
2
Western Blot Analysis of Cell Lysates
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Lysates from cells lysed in cold RIPA buffer supplemented with protease inhibitors were subjected to standard Western Blot protocols (8 (link)). The primary antibodies were α-GAPDH (glyceraldehyde-3-phosphate dehydrogenase; 1:10,000), α-E-cadherin (1:1,000), α-ST6GALNAC5 (1:1,000), and α-Vimentin (1:10,000) from Abcam and α-B3GLCT (B3GALTL, 1:1,000) and α-ST3GAL5 (1:1,000) from Novus Biologicals. Secondary antibodies [α-mouse-HRP (GAPDH) or α-rabbit-HRP (all others) (Bio-Rad)] were used at 1:5,000. Blots were developed using SuperSignal West Pico (Thermo Scientific).
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