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Bob nepx

Manufactured by Addgene
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BOB-NepX is a laboratory equipment product. It is a device used for the purification and enrichment of target proteins or biomolecules from complex samples. The core function of BOB-NepX is to facilitate the separation and isolation of specific molecules of interest through affinity-based capture and elution.

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Lab products found in correlation

Optima le 80k, by Beckman Coulter (2 mentions) Lipofectamine 3000, by Thermo Fisher Scientific (2 mentions) Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (2 mentions) Pspax2, by Addgene (2 mentions) Hyclone cosmic calf serum, by GE Healthcare (2 mentions) Pmd2 g, by Addgene (2 mentions) Plko 1, by Addgene (2 mentions)

2 protocols using bob nepx

1

Lentiviral Transduction of Mouse Cortical Neurons

Check if the same lab product or an alternative is used in the 5 most similar protocols
Lentiviral particles for shRNA knockdowns and hTau 2N4R and 0N4R protein expression in mouse cortical neurons were prepared as follows. The expression plasmids, pCSC-SP-PW-NepX (also known as BOB-NepX) or pLKO.1 (Addgene plasmids 12,340 and 10,878, respectively), and the packaging vectors, pSPAX2 and pMD2.G (Addgene plasmids 12,260 and 12,259, respectively) were transfected using Lipofectamine 3000 (ThermoFisher) into HEK293T cells grown in 15 cm Petri dishes to ~80% confluence in DMEM (GIBCO) supplemented with 10% HyClone cosmic calf serum (GE Healthcare). Each transfection was with 15 μg total DNA at a 50%/37.5%/12.5% ratio of expression vector/pSPAX2/pMD2.G. Lentivirus-conditioned medium was collected 24 and 48 h after the start of transfection. Lentiviral particles were concentrated in a Beckman Coulter Optima LE-80 K ultracentrifuge for 2 h at 23,000 rpm (95,000 gav) at 4 °C in an SW28 rotor, resuspended in 400 μl Neurobasal medium and stored at −80 °C in 20 μl aliquots. Cultured neurons were transduced in Neurobasal/B27 medium and incubated for 72 h before assays were performed.
Norambuena A., Sun X., Wallrabe H., Cao R., Sun N., Pardo E., Shivange N., Wang D.B., Post L.A., Ferris H.A., Hu S., Periasamy A, & Bloom G.S. (2022). SOD1 mediates lysosome-to-mitochondria communication and its dysregulation by amyloid-β oligomers. Neurobiology of disease, 169, 105737.
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2

Lentiviral Transduction of Mouse Cortical Neurons

Check if the same lab product or an alternative is used in the 5 most similar protocols
Lentiviral particles for shRNA knockdowns and hTau 2N4R and 0N4R protein expression in mouse cortical neurons were prepared as follows. The expression plasmids, pCSC-SP-PW-NepX (also known as BOB-NepX) or pLKO.1 (Addgene plasmids 12,340 and 10,878, respectively), and the packaging vectors, pSPAX2 and pMD2.G (Addgene plasmids 12,260 and 12,259, respectively) were transfected using Lipofectamine 3000 (ThermoFisher) into HEK293T cells grown in 15 cm Petri dishes to ~80% confluence in DMEM (GIBCO) supplemented with 10% HyClone cosmic calf serum (GE Healthcare). Each transfection was with 15 μg total DNA at a 50%/37.5%/12.5% ratio of expression vector/pSPAX2/pMD2.G. Lentivirus-conditioned medium was collected 24 and 48 h after the start of transfection. Lentiviral particles were concentrated in a Beckman Coulter Optima LE-80 K ultracentrifuge for 2 h at 23,000 rpm (95,000 gav) at 4 °C in an SW28 rotor, resuspended in 400 μl Neurobasal medium and stored at −80 °C in 20 μl aliquots. Cultured neurons were transduced in Neurobasal/B27 medium and incubated for 72 h before assays were performed.
Norambuena A., Sun X., Wallrabe H., Cao R., Sun N., Pardo E., Shivange N., Wang D.B., Post L.A., Ferris H.A., Hu S., Periasamy A, & Bloom G.S. (2022). SOD1 mediates lysosome-to-mitochondria communication and its dysregulation by amyloid-β oligomers. Neurobiology of disease, 169, 105737.
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