Detects human, mouse, rat Nucleostemin in direct ELISAs and Western blots. In direct ELISAs, less than 1% cross-reactivity with recombinant rat Nucleostemin C-terminal peptide (aa 281-538) is observed. Rat Nucleostemin specific IgG was purified by first passing the sera over a rat Nucleostemin aa 2-538 column and then passing the bound fraction over a rat Nucleostemin aa 281-538 column to removed rat Nucleostemin C-terminal specific IgG.
Polyclonal Goat IgG
E. coli-derived recombinant rat Nucleostemin Lys2-Ile538 Accession # Q811S9
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Detection of Human Nucleostemin by Western Blot. Western blot shows lysates of PC‑3 human prostate cancer cell line and SW480 human colorectal adenocarcinoma cell line. PVDF membrane was probed with 0.25 µg/mL of Goat Anti-Human/Mouse/Rat Nucleostemin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1638) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). A specific band was detected for Nucleostemin at approximately 70 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Nucleostemin in U2OS Human Cell Line. Nucleostemin was detected in immersion fixed U2OS human osteosarcoma cell line using 10 µg/mL Goat Anti-Human/Mouse/Rat Nucleostemin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1638) for 3 hours at room temperature. Cells were stained with the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red, upper panel; Catalog # NL001) and counterstained with DAPI (blue, lower panel). View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Detection of Human Nucleostemin by Simple WesternTM. Simple Western lane view shows lysates of PC‑3 human prostate cancer cell line, loaded at 0.2 mg/mL. A specific band was detected for Nucleostemin at approximately 83 kDa (as indicated) using 5 µg/mL of Goat Anti-Human/Mouse/Rat Nucleostemin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1638) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.
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.
Nucleostemin is a protein found in the nucleoli of embryonic stem cells, adult CNS stem cells, primitive cells in the bone marrow and cancer cells. It is not in the differentiated cells of most adult tissues. It has been suggested to play a role in controlling the cell-cycle progression in stem cells and cancer cells (1-3).
Tsai, R.Y. and R.D. McKay (2002) Genes Dev. 16:2991.
Baddoo, M. et al. (2003) J. Cell Biochem. 89:1235.
R&D Systems personnel manually curate a database that contains references using R&D Systems products.
The data collected includes not only links to publications in PubMed,
but also provides information about sample types, species, and experimental conditions.
The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.