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Anti-Nup133, clone 9C2H8, Cat. No. MABE1926, is a rat monoclonal antibody that detects Nuclear pore complex protein Nup133 and is tested for use in Immunofluorescence and Western Blotting.
More>>Anti-Nup133, clone 9C2H8, Cat. No. MABE1926, is a rat monoclonal antibody that detects Nuclear pore complex protein Nup133 and is tested for use in Immunofluorescence and Western Blotting. Less<<
Empfohlene Produkte
Übersicht
Replacement Information
Description
Catalogue Number
MABE1926-100UL
Description
Anti-Nup133 Antibody, clone 9C2H8
Alternate Names
Nuclear pore complex protein Nup133
133 kDa nucleoporin
Nucleoporin Nup133
Background Information
Nuclear pore complex protein Nup133 (UniProt: Q8R0G9; also known as 133 kDa nucleoporin, Nucleoporin Nup133) is encoded by the Nup133 gene (Gene ID: 234865) in murine species. Trafficking between the nucleus and the cytoplasm occurs via the nuclear pore complexes (NPCs), which are large supramolecular assemblies of ~120 MDa embedded in the double-membrane nuclear envelope. They are composed of 34 distinct protein subunits, termed nucleoporins (Nups), each present in multiple copies, leading to an assembly of more than 1000 polypeptides. Nup133 is a part of the Nup107-160 subcomplex (also termed Y-complex or coat nucleoporin complex, CNC) that is composed of Nup160, Nup133, Nup107, Nup96, Nup85, Nup43, Nup37, Seh1, Sec13, and ELYS in vertebrates. This complex is located on both the cytoplasmic and nuclear side of the nuclear pore in interphase and at kinetochores during mitosis. The N-terminal domain of Nup133 folds into a seven-bladed beta propeller structure and its C-terminal domain anchors it to the subcomplex via Nup107. It has been reported that mouse Nup133, although required for embryonic development beyond gastrulation stages, is dispensable for embryonic stem cell proliferation and is not essential for assembly of NPC scaffold in these pluripotent cells. (Ref.: Belgareh, N., et al., 2001, Cell Biol. 154(6); 1147-1160; Berke, IC., et al. (2004). J. Cell Biol. 167(4); 591-597; Lupu, F., et al. (2008) Dev Cell. 14(6); 831-842; Souquet, B., et al. (2018). Cell Rep. 23(8); 2443-2454; von Apen, A., and Beck, M., (2016). J. Mol. Biol. 428 (10 Pt A); 2001-2110).
References
Product Information
Format
Purified
Presentation
Purified rat monoclonal antibody IgG2a in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide.
Anti-Nup133, clone 9C2H8, Cat. No. MABE1926, is a rat monoclonal antibody that detects Nuclear pore complex protein Nup133 and is tested for use in Immunofluorescence and Western Blotting.
Key Applications
Western Blotting
Immunofluorescence
Application Notes
Immunofluorescence Analysis: A representative lot detected Nup133 in Immunofluorescence applications (Berto, A., et. al. (2018). EMBO Rep. 19(5); Souquet, B., et. al. (2018). Cell Rep. 23(8):2443-2454).
Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user
Biological Information
Immunogen
His-tagged recombinant fragment corresponding to 447 amino acids from the N-terminal half of murine Nuclear pore complex protein Nup133.
Epitope
N-terminal half
Clone
9C2H8
Concentration
0.5 mg/mL. Please refer to guidance on suggested starting dilutions and/or titers per application and sample type.
Host
Rat
Specificity
Clone 9C2H8 is a rat monoclonal antibody that detects murine Nuclear pore complex protein Nup133. It targets an epitope within the N-terminal half.
~130 kDa observed; 128.62 kDa calculated. Uncharacterized bands may be observed in some lysate(s).
Physicochemical Information
Dimensions
Materials Information
Toxicological Information
Safety Information according to GHS
Safety Information
Product Usage Statements
Quality Assurance
Evaluated by Western Blotting in Mouse embryonic stem cell (ESC) lysates.
Western Blotting Analysis: A 1:500 dilution of this antibody detected Nup133 in Mouse embryonic stem cell (ESC) lysates.
Usage Statement
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
Storage and Shipping Information
Storage Conditions
Stable for 1 year at +2°C to +8°C from date of receipt.
Nup133 Is Required for Proper Nuclear Pore Basket Assembly and Dynamics in Embryonic Stem Cells Nup133 belongs to the Y-complex, a key component of the nuclear pore complex (NPC) scaffold. Studies on a null mutation in mice previously revealed that Nup133 is essential for embryonic development but not for mouse embryonic stem cell (mESC) proliferation. Using single-pore detection and average NE-fluorescence intensity, we find that Nup133 is dispensable for interphase and postmitotic NPC scaffold assembly in pluripotent mESCs. However, loss of Nup133 specifically perturbs the formation of the nuclear basket as manifested by the absence of Tpr in about half of the NPCs combined with altered dynamics of Nup153. We further demonstrate that its central domain mediates Nup133's role in assembling Tpr and Nup153 into a properly configured nuclear basket. Our findings thus revisit the role of the Y-complex in pore biogenesis and provide insights into the interplay between NPC scaffold architecture, nuclear basket assembly, and the generation of heterogeneity among NPCs. Cell Rep
23(8)
2443-2454
2018
Nup133 belongs to the Y-complex, a key component of the nuclear pore complex (NPC) scaffold. Studies on a null mutation in mice previously revealed that Nup133 is essential for embryonic development but not for mouse embryonic stem cell (mESC) proliferation. Using single-pore detection and average NE-fluorescence intensity, we find that Nup133 is dispensable for interphase and postmitotic NPC scaffold assembly in pluripotent mESCs. However, loss of Nup133 specifically perturbs the formation of the nuclear basket as manifested by the absence of Tpr in about half of the NPCs combined with altered dynamics of Nup153. We further demonstrate that its central domain mediates Nup133's role in assembling Tpr and Nup153 into a properly configured nuclear basket. Our findings thus revisit the role of the Y-complex in pore biogenesis and provide insights into the interplay between NPC scaffold architecture, nuclear basket assembly, and the generation of heterogeneity among NPCs.
Cenp-F is a multifaceted protein implicated in cancer and developmental pathologies. The Cenp-F C-terminal region contains overlapping binding sites for numerous proteins that contribute to its functions throughout the cell cycle. Here, we focus on the nuclear pore protein Nup133 that interacts with Cenp-F both at nuclear pores in prophase and at kinetochores in mitosis, and on the kinase Bub1, known to contribute to Cenp-F targeting to kinetochores. By combining in silico structural modeling and yeast two-hybrid assays, we generate an interaction model between a conserved helix within the Nup133 β-propeller and a short leucine zipper-containing dimeric segment of Cenp-F. We thereby create mutants affecting the Nup133/Cenp-F interface and show that they prevent Cenp-F localization to the nuclear envelope, but not to kinetochores. Conversely, a point mutation within an adjacent leucine zipper affecting the kinetochore targeting of Cenp-F KT-core domain impairs its interaction with Bub1, but not with Nup133, identifying Bub1 as the direct KT-core binding partner of Cenp-F. Finally, we show that Cenp-E redundantly contributes together with Bub1 to the recruitment of Cenp-F to kinetochores.
