Cells were extracted using radioimmunoprecipitation assay (RIPA) lysis buffer (50 mmol/l Tris-HCl pH 7.4, 150 mmol/l NaCl, 1% Triton X-100, 1% sodium deoxycholate, 0.1% SDS, 1 mmol/l EDTA, protease inhibitors) and centrifuged for 15 min at 12.000 g. 30 μg protein were separated by SDS-PAGE and blotted onto Roti®Fluoro PVDF membrane (Roth, Karlsruhe, Germany). Membranes were incubated for 1 h at room temperature with the following primary antibodies: anti-c-Myc (1:1,000) and anti-GAPDH (1:1,000) (Santa-Cruz Biotechnology, Santa Cruz, CA, USA) or anti-MiD51 (1:1,000) (Proteintech, Rosemont, IL, USA) and anti-beta-actin (1:200) (Santa Cruz). Immunoreactive bands were visualized using the following fluorescence-labeled secondary antibodies: IRDye 680 CW, IRDye 800 CW and analyzed via the Odyssey imaging system. Densitometry measurements of bands were performed using the Odyssey infrared imaging system (LI-COR, Lincoln, NE, USA).
Roti fluoro pvdf membrane
Manufactured by Carl Roth
Sourced in Germany
Roti®Fluoro PVDF membrane is a polyvinylidene fluoride (PVDF) membrane designed for use in various laboratory applications. It is a durable and chemically resistant material that can withstand a wide range of solvents and pH conditions. The membrane is suitable for techniques such as Western blotting, dot blotting, and protein transfer.
Automatically generated - may contain errors
Lab products found in correlation
Anti gapdh, by Santa Cruz Biotechnology (2 mentions)
Odyssey infrared imaging system, by LI COR (2 mentions)
Anti c myc, by Santa Cruz Biotechnology (1 mentions)
Anti β actin, by Santa Cruz Biotechnology (1 mentions)
Anti mid51, by Proteintech (1 mentions)
Anti fis1, by Santa Cruz Biotechnology (1 mentions)
Anti β tubulin, by Santa Cruz Biotechnology (1 mentions)
Mitoprofile total oxphos rodent wb antibody cocktail, by Abcam (1 mentions)
2 protocols using roti fluoro pvdf membrane
1
Western Blot Analysis of c-Myc and MiD51
2
SDS-PAGE and Western Blot Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Cells were extracted with RIPA buffer (50 mmol/L Tris-HCl pH 7.4, 150 mmol/L NaCl, 1% Triton X-100, 1% sodium deoxycholate, 0.1% SDS, 1 mmol/L EDTA, protease inhibitors) and centrifuged for 10 min at 10,000 × g. Forty micrograms of proteins were separated by SDS-PAGE and blotted onto Roti Fluoro PVDF membrane (Roth, Karlsruhe, Germany). The membranes were probed for 1 h at room temperature with the following primary antibodies: anti-beta-tubulin (1:1000), anti-FIS1 (1:1000), anti-GAPDH (1:1000) (Santa-Cruz Biotechnology) and the MitoProfile Total OXPHOS Rodent WB Antibody Cocktail (MitoSciences, Eugene, OR, USA). Immunoreactive bands were visualised using the following fluorescence-labelled secondary antibodies: IRDye 680, IRDye 800 CW and analysed via the Odyssey imaging system. Densitometry measurements of bands were performed using the Odyssey infrared imaging system (LI-COR, Lincoln, NE, USA) and normalised to beta-tubulin or GAPDH expression.
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