Nucleoporin NUP85 in Human MSCs. Nucleoporin NUP85 was detected in immersion fixed human multipotent stromal cells (MSCs) using Sheep Anti-Human Nucleoporin NUP85 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5976) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 493-conjugated Anti-Sheep IgG Secondary Antibody (green; Catalog # NL012) and counterstained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Detection of Human Nucleoporin NUP85 by Western Blot. Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line. PVDF membrane was probed with 1 µg/mL of Sheep Anti-Human Nucleoporin NUP85 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5976) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for Nucleoporin NUP85 at approximately 70 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 8.
Preparation and Storage
Reconstitute at 0.2 mg/mL in sterile PBS.
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: Nucleoporin NUP85
NUP85 (Nucleoporin 85; also Nup75/nucleoporin 75 and FROUNT) is a 70‑72 kDa member of the nucleoporin Nup85 family of proteins. It is one of nine members of the Nup107-160 subcomplex that performs at least two functions in the cell. First, it serves as an assembly point for nuclear pore formation, and may stabilize the bend that normally exists between the inner and outer nuclear membranes. Second, it appears to be quite stable, and during mitosis, promotes mitotic spindle assembly. Human NUP85 is 656 amino acids (aa) in length. It is an alpha -helical protein with no obvious domain(s) or structural motif. Phosphorylation on Ser233, however, may regulate the overall interaction of the NUP107‑160 subcomplex with the much larger nuclear pore. There one potential splice form that shows an alternative start site at Met47. Full-length NUP85 shares 92% aa identity with mouse NUP85.
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