Overview

  • Product name
    Anti-Nucleostemin antibody
    See all Nucleostemin primary antibodies
  • Description
    Rabbit polyclonal to Nucleostemin
  • Tested applications
    Suitable for: ICC/IF, IHC-P, WB, IPmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Chicken, Human
    Predicted to work with: Rabbit, Horse, Dog, Chimpanzee
  • Immunogen

    Synthetic peptide corresponding to a region between residue 500 and the C-terminus (residue 549) of human Nucleostemin (NP_055181.3)

  • Positive control
    • HeLa, 293T and mouse 3T3 whole cell lysates.

Properties

Applications

Our Abpromise guarantee covers the use of ab70346 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
ICC/IF Use a concentration of 1 µg/ml.
IHC-P 1/200 - 1/1000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
WB 1/2000 - 1/10000. Detects a band of approximately 62 kDa (predicted molecular weight: 62 kDa).
IP Use at 1-4 µg/mg of lysate.

Target

  • Function
    May be required to maintain the proliferative capacity of stem cells and may play an important role in tumorigenesis.
  • Tissue specificity
    Increased levels in lung tissue in cancer patients.
  • Sequence similarities
    Belongs to the MMR1/HSR1 GTP-binding protein family.
    Contains 1 G (guanine nucleotide-binding) domain.
  • Domain
    The basic domain (B) allows nucleolar localization in the absence of GTP. The intermediate domain (I) inhibits nucleolar localization by the B domain and is required for exit from the nucleolus. Exit from the nucleolus to the nucleoplasm requires both the I and the acidic (A) domains, and may be triggered by GTP hydrolysis.
    In contrast to other GTP-binding proteins, this family is characterized by a circular permutation of the GTPase motifs described by a G4-G1-G3 pattern.
  • Cellular localization
    Nucleus. Nucleus > nucleolus. Shuttles between the nucleus and nucleolus.
  • Information by UniProt
  • Database links
  • Alternative names
    • C77032 antibody
    • E2 induced gene 3 protein antibody
    • E2-induced gene 3 protein antibody
    • E2IG3 antibody
    • Estradiol induced nucleotide binding protein antibody
    • GNL3 antibody
    • GNL3_HUMAN antibody
    • Guanine nucleotide binding protein like 3 (nucleolar) antibody
    • Guanine nucleotide binding protein like 3 antibody
    • Guanine nucleotide-binding protein-like 3 antibody
    • MGC800 antibody
    • NNP47 antibody
    • Novel nucleolar protein 47 antibody
    • NS antibody
    • Nucleolar GTP binding protein 3 antibody
    • Nucleolar GTP-binding protein 3 antibody
    • Nucleostemin antibody
    • Putative nucleotide binding protein estradiol induced antibody
    see all

Anti-Nucleostemin antibody images

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human skin carcinoma tissue labelling Nucleostemin with ab70346 at 1/1000 (0.2µg/ml). Detection: DAB.
  • All lanes : Anti-Nucleostemin antibody (ab70346) at 0.04 µg/ml

    Lane 1 : HeLa whole cell lysate at 50 µg
    Lane 2 : HeLa whole cell lysate at 15 µg
    Lane 3 : HeLa whole cell lysate at 5 µg
    Lane 4 : 293T whole cell lysate at 50 µg
    Lane 5 : mouse 3T3 whole cell lysate at 50 µg


    Predicted band size : 62 kDa
    Observed band size : 62 kDa


    Exposure time : 10 seconds
  • Detection of Nucleostemin by Western Blot of Immunprecipitate. All lanes: ab70346 at 1µg/ml staining Nucleostemin in HeLa whole cell lysates immunoprecipitated at 3µg/mg lysate (1 mg/IP; 20% of IP loaded/lane) using the following antibodies: Lane 1: an irrelevant antibody. Lane 2: ab70345 Lane 3: ab70346. Detection: Chemiluminescence with exposure time of 3 seconds
  • Immunohistochemistical detection of Nucleostemin with ab70346 on formaldehyde-fixed paraffin-embedded human colon sections. Antigen retrieval step: heat mediated in citric acid pH6 buffer. Blocking step: 1% BSA  for 10 mins @ rt°C. Primary antibody ab70346 incubated at 1/50 for 16 hours in TBS/BSA/azide. Secondary antibody: anti-rabbit IgG conjugated to biotin (1/200).  The intestinal epithelium is the most rapidly self-renewing tissue in adult mammals. Stem cell markers have been observed in cycling columnar cells at the crypt base (Nature 449, 1003-1007; 2007). The cells (nuclei of the simple columnar epithelium of the upper mucosa) immunoreactive for Nucleostemin in this image (green arrowheads) anatomically correspond to the same cells expected to be Lgr5 positivite. A red arrowhead indicates nucleolar positivity in one of many cell nuclei in the Lamina propria of the mucosa (this area is not known for stem cell posit

    See Abreview

  • ICC/IF image of ab70346 stained HeLa cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab70346, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

References for Anti-Nucleostemin antibody (ab70346)

This product has been referenced in:
  • Allen KD  et al. Nucleolar integrity is required for the maintenance of long-term synaptic plasticity. PLoS One 9:e104364 (2014). ICC/IF ; Mouse . Read more (PubMed: 25089620) »

See 1 Publication for this product

Product Wall

Application
Western blot
Sample
Human Cell lysate - whole cell (U2OS)
Gel Running Conditions
Reduced Denaturing
Loading amount
25 µg
Specification
U2OS
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
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Abcam user community

Verified customer

Submitted Sep 12 2017

Application
Western blot
Loading amount
20 µg
Gel Running Conditions
Reduced Denaturing (10 % GEL, SDS)
Sample
Human Cell lysate - whole cell (HEK293, HELA, HCT116)
Specification
HEK293, HELA, HCT116
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Username

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Verified customer

Submitted Apr 24 2014

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
Serum as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 21°C
Sample
Human Cell (HEK293)
Specification
HEK293
Permeabilization
Yes - TRITON
Fixative
Methanol
Username

Abcam user community

Verified customer

Submitted Apr 23 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (Hela Cells)
Specification
Hela Cells
Fixative
Paraformaldehyde
Permeabilization
Yes - .25% Triton X-100 in PBS
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
Username

Ms. Western,IP ChIP

Verified customer

Submitted Oct 22 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Tissue lysate - whole (Hela Cells)
Loading amount
30 µg
Specification
Hela Cells
Gel Running Conditions
Reduced Denaturing (10% SDS-PAGE gel)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: RT°C
Username

Abcam user community

Verified customer

Submitted Oct 22 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Tissue lysate - whole (Mouse ES cells)
Loading amount
30 µg
Specification
Mouse ES cells
Gel Running Conditions
Reduced Denaturing (10% SDS-PAGE gel)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: RT°C
Username

Abcam user community

Verified customer

Submitted Oct 22 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunoprecipitation
Sample
Human Cell lysate - whole cell (Hela Cells)
Total protein in input
200000 µg
Specification
Hela Cells
Immuno-precipitation step
Protein G
Username

Abcam user community

Verified customer

Submitted Oct 21 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Colon)
Specification
Colon
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid pH6
Permeabilization
No
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: rt°C
Username

Mr. Carl Hobbs

Verified customer

Submitted Feb 15 2010

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Chicken Tissue sections (chick day 6 embryo liver and kidney)
Specification
chick day 6 embryo liver and kidney
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid pH6
Permeabilization
No
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 1%
Username

Mr. Carl Hobbs

Verified customer

Submitted Feb 15 2010

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rat Tissue sections (adult rat brain)
Specification
adult rat brain
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: citric acid pH6
Permeabilization
No
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: rt°C
Username

Mr. Carl Hobbs

Verified customer

Submitted Feb 15 2010

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