Cells were placed on coverslips in 24-well plates and then washed with cold PBS three times. Then, the coverslips were fixed with 4% formaldehyde and immunostained using monoclonal antibodies of E-cadherin and Vimentin. After that, DNA of these cells was stained with 4′,6-diamidino-2-phenylindole (DAPI). The Alexa Fluor 546 (red) goat anti-rabbit antibody (Molecular Probes, OR, USA) was used for E-cadherin (no. 14472, 1:50; Cell Signaling Technology, Boston, MA, USA) and Vimentin (sc-66002, 1:50; Santa Cruz Biotechnology, Santa Cruz, CA, USA). Immunofluorescence was determined using confocal microscopy (Leica TCS SP5).
Sc 66002
Manufactured by Santa Cruz Biotechnology
Sourced in United States
Sc-66002 is a laboratory instrument designed for specialized research applications. It is a high-precision device that serves as a core component for various experimental procedures. The product specifications and technical details are available upon request.
Automatically generated - may contain errors
Lab products found in correlation
Tcs sp5, by Leica (1 mentions)
Alexa fluor 546 red goat anti rabbit antibody, by Thermo Fisher Scientific (1 mentions)
Polyvinylidene difluoride membrane, by Bio-Rad (1 mentions)
Ecl prime, by GE Healthcare (1 mentions)
β actin, by Merck Group (1 mentions)
Horseradish peroxidase conjugated anti mouse or anti rabbit antibodies, by Bio-Rad (1 mentions)
2 protocols using sc 66002
1
Immunostaining of Cell Adhesion Proteins
2
Western Blot Analysis of Vimentin and AMBP
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Protein samples (20 μg) were separated by 10% sodium dodecyl sulfate−polyacrylamide gel electrophoresis under reducing conditions and transferred to a polyvinylidene difluoride membrane (Bio-Rad, Hemel Hempstead, UK). After incubation with 5% nonfat milk in a mixture of Tris-buffered saline and Tween 20, the membrane was incubated with anti-vimentin (SC-66002, 1:500, Santa Cruz Biotechnology, Santa Cruz, CA) or anti−α-1-microglobulin/bikunin precursor (anti-AMBP) (SC-81948, 1:400, Santa Cruz Biotechnology, Santa Cruz, CA) antibody at 4°C for 12 hours. β-Actin was used as a loading control (1:250, Sigma-Aldrich). Membranes were then incubated with horseradish peroxidase-conjugated anti-mouse or anti-rabbit antibodies (1:50000, Bio-Rad) and developed with ECL Prime (GE Healthcare, Little Chalfont, UK). Detection was performed with the x-ray film Agfa CP-BU new (Agfa Healthcare, Mortsel, Belgium).
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