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Recombinant smn

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Recombinant SMN is a laboratory product that provides the survival motor neuron (SMN) protein. The SMN protein is a key component involved in the assembly and function of small nuclear ribonucleoproteins (snRNPs), which are essential for pre-mRNA splicing. This product can be used for research purposes related to the study of SMN and its role in cellular processes.

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Lab products found in correlation

Tla100.2 rotor, by Beckman Coulter (2 mentions) Sample buffer, by Santa Cruz Biotechnology (2 mentions)

2 protocols using recombinant smn

1

Purification and Characterization of SMN-Bound Ribosomes

Cited 3 times
Check if the same lab product or an alternative is used in the 5 most similar protocols
Purification of 80S ribosomes was performed from NSC-34 depleted from SMN using CRISPR-Cas9 technology (11 (link)). The lysate was treated with RNase I (7.5 Units /1 Abs260 lysate) at RT for 45 min and analyzed by sucrose gradient (10-40%) (65 (link)). The fraction corresponding to the 80S was collected and 2mM DTT was added. After centrifugation at 90,000 rpm for 4 h using a TLA100.2 rotor (Beckmann) the pellet was resuspended in 10 mM Tris-HCl pH 7.5, 10 mM MgCl2, 150 mM NaCl, 2 mM DTT, 100 μg/mL cycloheximide and stored at −80°C. Ribosome concentration was calculated as in (66 (link)). Recombinant SMN was purchased (ENZO) and incubated with ribosomes for 2 h at 4°C. SMN bound ribosomes were purified from unbound SMN by ultracentrifugation at 90,000 rpm for 4 h using the TLA100.2 rotor on 30% sucrose cushion. The supernatant was kept for protein purification by using the chloroform/methanol protocol and the pellet was directly dissolved in sample buffer (Santa Cruz), heated at 99°C for 10 min and resolved by SDS-PAGE. SMN and RPL26, were detected using primary and secondary antibodies described above.
Lauria F., Bernabò P., Tebaldi T., Groen E.J., Perenthaler E., Maniscalco F., Rossi A., Donzel D., Clamer M., Marchioretto M., Omersa N., Orri J., Serra M.D., Anderluh G., Quattrone A., Inga A., Gillingwater T.H, & Viero G. (2020). SMN-primed ribosomes modulate the translation of transcripts related to Spinal Muscular Atrophy. Nature cell biology, 22(10), 1239-1251.
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2

Purification and Characterization of SMN-Bound Ribosomes

Cited 3 times
Check if the same lab product or an alternative is used in the 5 most similar protocols
Purification of 80S ribosomes was performed from NSC-34 depleted from SMN using CRISPR-Cas9 technology (11 (link)). The lysate was treated with RNase I (7.5 Units /1 Abs260 lysate) at RT for 45 min and analyzed by sucrose gradient (10-40%) (65 (link)). The fraction corresponding to the 80S was collected and 2mM DTT was added. After centrifugation at 90,000 rpm for 4 h using a TLA100.2 rotor (Beckmann) the pellet was resuspended in 10 mM Tris-HCl pH 7.5, 10 mM MgCl2, 150 mM NaCl, 2 mM DTT, 100 μg/mL cycloheximide and stored at −80°C. Ribosome concentration was calculated as in (66 (link)). Recombinant SMN was purchased (ENZO) and incubated with ribosomes for 2 h at 4°C. SMN bound ribosomes were purified from unbound SMN by ultracentrifugation at 90,000 rpm for 4 h using the TLA100.2 rotor on 30% sucrose cushion. The supernatant was kept for protein purification by using the chloroform/methanol protocol and the pellet was directly dissolved in sample buffer (Santa Cruz), heated at 99°C for 10 min and resolved by SDS-PAGE. SMN and RPL26, were detected using primary and secondary antibodies described above.
Lauria F., Bernabò P., Tebaldi T., Groen E.J., Perenthaler E., Maniscalco F., Rossi A., Donzel D., Clamer M., Marchioretto M., Omersa N., Orri J., Serra M.D., Anderluh G., Quattrone A., Inga A., Gillingwater T.H, & Viero G. (2020). SMN-primed ribosomes modulate the translation of transcripts related to Spinal Muscular Atrophy. Nature cell biology, 22(10), 1239-1251.
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