Nup133

Manufactured by Santa Cruz Biotechnology

Sourced in United States

NUP133 is a nuclear pore complex protein that is involved in the formation and maintenance of the nuclear envelope. It plays a crucial role in the transport of molecules between the nucleus and the cytoplasm.

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3 protocols using nup133

Trim21-Mediated Nups Knockdown in iPSNs

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Trim21 mediated knockdown of Nups was modified from a previously described protocol (Clift et al., 2017 (link)). POM121 (Thermo Fisher Scientific), Nup133 (Santa Cruz Biotechnology), NDC1 (Novus Biologicals), and GAPDH (Cell Signaling) antibodies were dialyzed using Slide-A-Lyzer MINI Dialysis Device, 20K MWCO (Thermo Fisher Scientific). Following dialysis, antibodies were concentrated using Amicon Ultra-0.5 Centrifugal Filter Unit with Ultracel-100 membrane (Millipore). Antibody concentration was determined using a NanoDrop 1000 spectrophotometer (Thermo Fisher Scientific) as described (Clift et al., 2017 (link)). On day 18 of differentiation, 5 × 10⁶ iPSNs were nucleofected with 5 μg of antibody and 4 μg Trim21 GFP plasmid DNA (Origene Technolgies) with Lonza P3 Primary Cell 4D Nucleofector Kit (Lonza) using program DC154. The next day, Trim21 GFP expression was observed and iPSNs were assayed on day 20 of differentiation.

Coyne A.N., Zaepfel B.L., Hayes L., Fitchman B., Salzberg Y., Luo E.C., Bowen K., Trost H., Aigner S., Rigo F., Yeo G.W., Harel A., Svendsen C.N., Sareen D, & Rothstein J.D. (2020). G4C2 repeat RNA initiates a POM121 mediated reduction in specific nucleoporins in C9orf72 ALS/FTD. Neuron, 107(6), 1124-1140.e11.

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Molecular Interactions in SV40 System

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SV40 large T antigen (Santa Cruz sc-147), monoclonal VP1 was provided by Dr. Walter Scott (University of Miami), polyclonal VP1 (Abcam ab53977), SV40 VP2/3 (Abcam ab53983), RanBP2 (Thermo Scientific A301-796A), Nesprin-2 (Bethyl A305-393A, Thermo Scientific MA5-18075), Hsp90 (Santa Cruz sc-13119), GFP (Dawen Cai, University of Michigan), FLAG (Millipore Sigma F7425), MAB414 NPC (Abcam ab24609), Histone H3 (Cell Signaling 9715S), BAP31 (Pierce MA3-002), Rab7 (Millipore Sigma R8779), Na+/K+ ATPase (Abcam ab76020), LAMP2 (Abcam ab18529), NUP35 (Millipore Sigma HPA018410), NUP88 (Santa Cruz sc-365868), NUP93 (Thermo Scientific A303-979A), NUP98 (Cell Signaling 2598S), NUP133 (Santa Cruz sc-376763), and NUP188 (Thermo Scientific PA5-66645), BICD2 (Abcam (ab117818).

Spriggs C.C., Cha G., Li J, & Tsai B. (2022). Components of the LINC and NPC complexes coordinately target and translocate a virus into the nucleus to promote infection. PLoS Pathogens, 18(9), e1010824.

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Comprehensive Antibody Characterization for Cell Imaging

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The following antibodies were used for Western blot (WB) and/or immunofluorescence (IF): NUP133 (sc-376763, Santa Cruz Biotechnology Inc., Dallas, TX, USA, WB 1:2000, IF 1:300), NUP107 (SAB2702098, Merck, WB 1:1000, IF 1:200), nuclear pore complex proteins mAb414 (ab24609, Abcam, Cambridge, UK, WB 1:1000, IF 1:200), beta-Actin (4970S, Cell Signaling Technology Inc., Danvers, MA, USA, IF 1:400), VCL (HPA063777, Atlas Antibodies AB, Bromma, Sweden, WB 1:1000), TUBA (T9026, Merck, WB 1:5000), FLAG M2 (F3165, Merck, WB 1:5000), PXN (610051, BD Biosciences, Franklin Lakes, NJ, USA, NU, 1:300), COL18A1 (ab275746, Abcam, WV 1:1000), PPME1 (sc-25278, Santa Cruz Biotechnology, WB 1:2000), Hoechst 33342, trihydrochloride, trihydrate (H3570, Thermo Fisher Scientific, IF 1:1000), and Alexa Fluor phalloidin 488/555 (Thermo Fisher Scientific, IF 1:200–1:750). For IF Alexa Fluor conjugated anti-mouse or anti-rabbit secondary antibodies (A-31572, A-21127, A21245, A31570, A28175, Thermo Fisher Scientific, IF 1:500) and WB HRP-linked, anti-mouse (goat anti-mouse immunoglobulins/HRP, P0447, Dako, WB 1:10,000) or anti-rabbit (goat anti-rabbit IgG, HRP-linked antibody, #7074, cell signaling, WB 1:1000) secondary antibodies were used.

Rogg M., Maier J.I., Ehle M., Sammarco A., Schilling O., Werner M, & Schell C. (2022). NUP133 Controls Nuclear Pore Assembly, Transcriptome Composition, and Cytoskeleton Regulation in Podocytes. Cells, 11(8), 1259.

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