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Human/Mouse/Rat Nucleostemin Antibody

Catalog #: AF1638
9 Citations Datasheet / COA / SDS
Catalog # Availability Size / Price Qty
AF1638
AF1638-SP
Detection of Human Nucleostemin by Western Blot.
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Citations (9)
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Human/Mouse/Rat Nucleostemin Antibody Summary

Species Reactivity
Human, Mouse, Rat
Specificity
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. 
Source
Polyclonal Goat IgG
Purification
Antigen Affinity-purified
Immunogen
E. coli-derived recombinant rat Nucleostemin
Lys2-Ile538
Accession # Q811S9
Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
Label
Unconjugated

Applications

Recommended Concentration
Sample
Western Blot
0.25 µg/mL
See below
Simple Western
5 µg/mL
See below
Immunocytochemistry
5-15 µg/mL
See below

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.

Scientific Data

Western Blot Detection of Human Nucleostemin antibody by Western Blot. View Larger

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.

Immunocytochemistry Nucleostemin antibody in U2OS Human Cell Line by Immunocytochemistry (ICC). View Larger

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.

Simple Western Detection of Human Nucleostemin antibody by Simple Western<sup>TM</sup>. View Larger

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.

Immunocytochemistry/ Immunofluorescence Detection of Human Nucleostemin by Immunocytochemistry/Immunofluorescence View Larger

Detection of Human Nucleostemin by Immunocytochemistry/Immunofluorescence Embryonic stem cell markers. Immunohistochemistry: A. NANOG; B. OCT4. The majority of nuclei in the NANOG IHC are brown indicating the presence of NANOG. Some, but not all, nuclei are positive for OCT4. Negative (secondary antibody only) and positive (seminoma) controls are presented in the thumbnails below the photomicrographs. C. Immunofluorescence against nucleostemin. Left: anti-nucleostemin antibody, center: DAPI, right: merge. Bar = 10 microns. Image collected and cropped by CiteAb from the following publication (https://bmcmolcellbiol.biomedcentral.com/articles/10.1186/1471-2121-15-20), licensed under a CC-BY license. Not internally tested by R&D Systems.

Reconstitution Calculator

Reconstitution Calculator

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.

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Preparation and Storage

Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS.
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Shipping
Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.
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: Nucleostemin

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).

References
  1. Tsai, R.Y. and R.D. McKay (2002) Genes Dev. 16:2991.
  2. Baddoo, M. et al. (2003) J. Cell Biochem. 89:1235.
  3. Normile, D. (2002) Science 298:1869.
Entrez Gene IDs
26354 (Human); 30877 (Mouse); 290556 (Rat)
Alternate Names
C77032; E2IG3NNP47; estradiol-induced nucleotide binding protein; GNL3; guanine nucleotide binding protein-like 3 (nucleolar); guanine nucleotide-binding protein-like 3; MGC800; Novel nucleolar protein 47; NSE2-induced gene 3 protein; Nucleolar GTP-binding protein 3; Nucleostemin

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Citations for Human/Mouse/Rat Nucleostemin Antibody

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.

9 Citations: Showing 1 - 9
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  1. Immunohistochemical study of doublecortin and nucleostemin in canine brain
    Authors: E. De Nevi, P. Marco-Salazar, D. Fondevila,1, E. Blasco, L. Pérez, M. Pumarola
    European Journal of Histochemistry
  2. Viral targeting of glioblastoma stem cells with patient-specific genetic and post-translational p53 deregulations
    Authors: J Gil-Ranedo, C Gallego-Ga, JM Almendral
    Cell Reports, 2021-09-07;36(10):109673.
    Species: Human
    Sample Types: Whole Cells
    Applications: ICC
  3. Overexpression of nucleostemin contributes to an advanced malignant phenotype and a poor prognosis in oral squamous cell carcinoma.
    Authors: Yoshida, R, Nakayama, H, Nagata, M, Hirosue, A, Tanaka, T, Kawahara, K, Nakagawa, Y, Matsuoka, Y, Sakata, J, Arita, H, Hiraki, A, Shinohara, M, Ito, T
    Br J Cancer, 2014-10-14;111(12):2308-15.
    Species: Human
    Sample Types: Cell Lysates, Whole Tissue
    Applications: IHC-P, Western Blot
  4. Phenotypic plasticity in normal breast derived epithelial cells.
    Authors: Sauder C, Koziel J, Choi M, Fox M, Grimes B, Badve S, Blosser R, Radovich M, Lam C, Vaughan M, Herbert B, Clare S
    BMC Cell Biol, 2014-06-10;15(0):20.
    Species: Human
    Sample Types: Whole Cells
    Applications: ICC
  5. Nucleostemin is indispensable for the maintenance and genetic stability of hematopoietic stem cells.
    Authors: Yamashita M, Nitta E, Nagamatsu G, Ikushima Y, Hosokawa K, Arai F, Suda T
    Biochem Biophys Res Commun, 2013-10-16;441(1):196-201.
    Species: Mouse
    Sample Types: Whole Cells
    Applications: IHC-Fr
  6. Reactive oxygen species regulate nucleostemin oligomerization and protein degradation.
    Authors: Huang M, Whang P, Chodaparambil JV
    J. Biol. Chem., 2011-01-17;286(13):11035-46.
    Species: Human
    Sample Types: Cell Lysates, Whole Cells
    Applications: ICC, Western Blot
  7. Depletion of guanine nucleotides leads to the Mdm2-dependent proteasomal degradation of nucleostemin.
    Authors: Huang M, Itahana K, Zhang Y, Mitchell BS
    Cancer Res., 2009-03-24;69(7):3004-12.
    Species: Human
    Sample Types: Cell Lysates, Whole Cells
    Applications: ICC, Western Blot
  8. Identification of Novel Markers That Demarcate the Nucleolus during Severe Stress and Chemotherapeutic Treatment
    Authors: Haitong Su, Mohamed Kodiha, Sunghoon Lee, Ursula Stochaj
    PLoS ONE
  9. Gold Nanoparticles Impinge on Nucleoli and the Stress Response in MCF7 Breast Cancer Cells
    Authors: Mohamed Kodiha, Hicham Mahboubi, Dusica Maysinger, Ursula Stochaj
    Nanobiomedicine (Rij)

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