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Sc 6243

Manufactured by Novus Biologicals
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Sc-6243 is a laboratory microplate reader designed for absorbance measurements. It is capable of reading microplates with 96 or 384 wells. The device uses a monochromator-based optical system to provide flexible wavelength selection between 200 nm and 999 nm.

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Lab products found in correlation

Lipofectamine 3000 transfection reagent, by Thermo Fisher Scientific (1 mentions) Fusion fx7 imager, by Vilber (1 mentions)

2 protocols using sc 6243

1

Studying NUPR1 and p53 SUMOylation in MiaPaCa-2 Cells

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MiaPaCa-2 cells were seeded on coverslips and transfected with 2 μg of plasmid DNA (Nupr1-Flag or importin-α3–Flag) using Lipofectamine 3000 Transfection Reagent (Thermo Fisher Scientific). MiaPaCa-2–6HF–SUMO1 cells, previously described (21 (link)), were used to measure the SUMOylation of TP53. At the end of the experiment, cells were washed in PBS, fixed, and permeabilized before immunostaining with Duolink In Situ (MilliporeSigma) following the manufacturer’s protocol. Anti-NUPR1 (rabbit, homemade), -p53 (rabbit, Santa Cruz Biotechnology, sc-6243), -MRE11 (rabbit, Novus Biologicals, NB100-142), or -Flag (mouse, MilliporeSigma, F1804) were used. Image acquisition was carried out on a Nikon Eclipse 90i fluorescence microscope. ImageJ (NIH) was used to count the number of red dots.
Lan W., Santofimia-Castaño P., Swayden M., Xia Y., Zhou Z., Audebert S., Camoin L., Huang C., Peng L., Jiménez-Alesanco A., Velázquez-Campoy A., Abián O., Lomberk G., Urrutia R., Rizzuti B., Geli V., Soubeyran P., Neira J.L, & Iovanna J. (2020). ZZW-115–dependent inhibition of NUPR1 nuclear translocation sensitizes cancer cells to genotoxic agents. JCI Insight, 5(18), e138117.
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2

Western Blot Analysis of Protein Targets

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Proteins were resolved by SDS-PAGE, and transferred to nitrocellulose membranes for 1 hour. Then, membranes were blocked 1 hour at room temperature with TBS (Tris buffered saline solution) and 5% BSA, and blotted overnight in TBS 5% BSA containing primary antibodies (1:500; anti-p53 [rabbit, Santa Cruz Biotechnology Inc., sc-6243], -MRE11 [rabbit, Novus Biologicals, NB100-142], -LSD1 [mouse, Santa Cruz Biotechnology Inc., sc-271720]). After extensive washes in TBS 0.1% Tween-20, membranes were incubated 1 hour at room temperature with HRP-conjugated secondary antibodies at 1:5000 before being revealed with Enhanced chemo-luminescence (ECL). Acquisition was performed with a Fusion FX7 imager (Vilber-Lourmat).
Lan W., Santofimia-Castaño P., Swayden M., Xia Y., Zhou Z., Audebert S., Camoin L., Huang C., Peng L., Jiménez-Alesanco A., Velázquez-Campoy A., Abián O., Lomberk G., Urrutia R., Rizzuti B., Geli V., Soubeyran P., Neira J.L, & Iovanna J. (2020). ZZW-115–dependent inhibition of NUPR1 nuclear translocation sensitizes cancer cells to genotoxic agents. JCI Insight, 5(18), e138117.
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