Anti-Nucleostemin antibody (ab70346)
- Datasheet
- Specific References (4)
- Protocols
Overview
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Product nameAnti-Nucleostemin antibody
See all Nucleostemin primary antibodies -
DescriptionRabbit polyclonal to Nucleostemin
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Host speciesRabbit
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Tested applicationsSuitable for: ICC/IF, IHC-P, WB, IPmore details
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Species reactivityReacts with: Mouse, Rat, Chicken, Human
Predicted to work with: Rabbit, Horse, Dog, Chimpanzee
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Immunogen
Synthetic peptide corresponding to a region between residue 500 and the C-terminus (residue 549) of human Nucleostemin (NP_055181.3)
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Positive control
- HeLa, 293T and mouse 3T3 whole cell lysates.
Properties
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FormLiquid
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Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
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Storage bufferPreservative: 0.09% Sodium Azide
Constituents: 0.1% BSA, Tris buffered saline -
Concentration information loading... -
PurityImmunogen affinity purified
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Purification notesab70346 was affinity purified using an epitope specific to Nucleostemin immobilized on solid support.
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ClonalityPolyclonal
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IsotypeIgG
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Research areas
Associated products
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Compatible Secondaries
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Positive Controls
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
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FunctionMay be required to maintain the proliferative capacity of stem cells and may play an important role in tumorigenesis.
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Tissue specificityIncreased levels in lung tissue in cancer patients.
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Sequence similaritiesBelongs to the MMR1/HSR1 GTP-binding protein family.
Contains 1 G (guanine nucleotide-binding) domain. -
DomainThe 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 localizationNucleus. Nucleus > nucleolus. Shuttles between the nucleus and nucleolus.
- Information by UniProt
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Database links
- Entrez Gene: 26354 Human
- Entrez Gene: 30877 Mouse
- Entrez Gene: 290556 Rat
- Omim: 608011 Human
- SwissProt: Q9BVP2 Human
- SwissProt: Q8CI11 Mouse
- SwissProt: Q811S9 Rat
- Unigene: 313544 Human
see all -
Alternative names
- C77032 antibody
- E2 induced gene 3 protein antibody
- E2-induced gene 3 protein antibody
see all
Images
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nucleostemin antibody (ab70346)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. -
Western blot - Anti-Nucleostemin antibody (ab70346)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 -
Immunoprecipitation - Anti-Nucleostemin antibody (ab70346)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 -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nucleostemin antibody (ab70346)This image is courtesy of an abreview submitted by Carl Hobbs, King's College London, United KingdomImmunohistochemistical 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 -
Immunocytochemistry/ Immunofluorescence - Anti-Nucleostemin antibody (ab70346)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.
Protocols
References
This product has been referenced in:
- Lu M et al. Dual-modal photoacoustic and magnetic resonance tracking of tendon stem cells with PLGA/iron oxide microparticles in vitro. PLoS One 13:e0193362 (2018). Read more (PubMed: 29608568) »
- Kim SJ et al. Effect of Bone Marrow Aspirate Concentrate-Platelet-Rich Plasma on Tendon-Derived Stem Cells and Rotator Cuff Tendon Tear. Cell Transplant 26:867-878 (2017). Read more (PubMed: 28105983) »
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