Anti-Nestin, clone 10C2
Recommended Replacement for: AB5922| Species Reactivity | Key Applications | Host | Format | Antibody Type |
|---|---|---|---|---|
| H , not M, not R | WB, IC, IH, IH(P) | Mouse | Purified | Monoclonal Antibody |
UniProt Number:
UniProt Summary
FUNCTION: SwissProt: P48681 # May play a role in the trafficking and distribution of IF proteins and potentially other cellular factors to daughter cells during progenitor cell division (By similarity).
SIZE: 1621 amino acids; 177439 Da
SUBUNIT: Interacts with FHOD3 (By similarity).
TISSUE SPECIFICITY: CNS stem cells.
DEVELOPMENTAL STAGE: Upon terminal neural differentiation, nestin is down-regulated and replaced by neurofilaments.
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SIZE: 1621 amino acids; 177439 Da
SUBUNIT: Interacts with FHOD3 (By similarity).
TISSUE SPECIFICITY: CNS stem cells.
DEVELOPMENTAL STAGE: Upon terminal neural differentiation, nestin is down-regulated and replaced by neurofilaments.
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Entrez Gene Number:
Entrez Gene Summary
Nestin is an intermediate filament protein that was first identified with a monoclonal antibody by Hockfield and McKay (1985) [PubMed 4078630]. It is expressed predominantly in stem cells of the central nervous system in the neural tube. Upon terminal neural differentiation, nestin is downregulated and replaced by neurofilaments.[supplied by OMIM]
Description:
Anti-Nestin, clone 10C2
Replaces:
AB5922
Background Information:
Nestin, a large intermediate filament protein (class Type VI) expressed during development and in myotendinous and neuromuscular junctions. Nestin expression is restricted, typically disappearing by E18. Nestin is thought to be a reasonable neuronal marker; however, recent studies have found nestin expression in other cell types such as endothelial cells {Folia Biologica, 44(5):155, 1998}. Nestin identifies the most primitive neuroepithelium but also identifies many other embryonic tissues, so it is not specific for CNS. Nestin expression is seen in almost all GBMs (Glioblastoma mulitformes) and many melanomas (both primary and metastatic) but not in any metastatic carcinoma.
Clone:
10C2
Specificity:
Reacts with human Nestin, with weak reactivity to monkey nestin. Not reactive with rodent nestin.
Key Applications:
- Western Blotting
- Immunocytochemistry
- Immunohistochemistry
- Immunohistochemistry (Paraffin)
Application Notes:
Immunoblotting: 1:1,000-1:2,000 on denatured protein. Western blot: 1:1000-1:5000. Recognizes a 220-240kDA band on denaturing SDS gels. There is also a 60-64kDa band present in most lysates, the identity of this smaller fragment is unknown but it is not seen in lysates not containing human nestin protein.
WI-38 (ATCC# CCL75) can be used as a positive control. U251 cell line is also positive.
Immunocytochemistry: 1:200 on cells fixed in 4% paraformaldehyde and permeabilized with 0.02% Triton
Immunohistochemistry: paraformaldehyde is the preferred fixative: 1:200 on formalin fixed, paraffin embedded tissue sections. Use triton 0.02% in block only; dilution buffer 0.1% BSA; incubations 1-2 hours RT. Human fetal brain is suggested as a positive control. (CA Messam et al, Develop. Brain Res, 134:87-92, 2002; Gu, et al., Brain Res. 943:174-180, 2002). Filamentous staining in the cytoplasm of cells
Optimal working dilutions must be determined by end user.
WI-38 (ATCC# CCL75) can be used as a positive control. U251 cell line is also positive.
Immunocytochemistry: 1:200 on cells fixed in 4% paraformaldehyde and permeabilized with 0.02% Triton
Immunohistochemistry: paraformaldehyde is the preferred fixative: 1:200 on formalin fixed, paraffin embedded tissue sections. Use triton 0.02% in block only; dilution buffer 0.1% BSA; incubations 1-2 hours RT. Human fetal brain is suggested as a positive control. (CA Messam et al, Develop. Brain Res, 134:87-92, 2002; Gu, et al., Brain Res. 943:174-180, 2002). Filamentous staining in the cytoplasm of cells
Optimal working dilutions must be determined by end user.
Species Reactivity:
Human
Non-Reactive Species:
- Mouse
- Rat
Isotype:
IgG1
Immunogen:
Fusion protein.
Epitope:
human
Molecular Weight:
200-220 kDa
Control:
Positive Control: WI-38 (ATCC# CCL75) or U251 cell lines
Presentation:
Protein A Purified mouse immunoglobulin in 20 mM sodium phosphate, 250 mM NaCl, pH. 7.6, with 0.1% sodium azide as a preservative.
Storage Conditions:
Antibodies may be stored at 4°C or -20°C. Avoid repeated freeze/thaw cycles.
During shipment, small volumes of product will occasionally become entrapped in the seal of the product vial. For products with volumes of 200µL or less, we recommend gently tapping the vial on a hard surface or briefly centrifuging the vial in a tabletop centrifuge to dislodge any liquid in the container's cap.
During shipment, small volumes of product will occasionally become entrapped in the seal of the product vial. For products with volumes of 200µL or less, we recommend gently tapping the vial on a hard surface or briefly centrifuging the vial in a tabletop centrifuge to dislodge any liquid in the container's cap.
Gene Symbol:
- NES
- ORFNames=Nbla00170
- FLJ21841
- Nestin.
- Nbla00170
Antibody Category:
Stem Cell Research
Antibody Sub-Category:
Neural Stem Cells
Usage Statement:
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
Concentration:
1 mg/mL
Stem Cell Workflow Stage:
Characterization
Stem Cell Type:
- Neural Stem Cells
- Epithelial Progenitor Cells
Format Code:
Pur
Trade Name:
Chemicon (Millipore)
Purification Method:
Protein A Purified
Format:
Purified
Antibody Type:
Monoclonal Antibody
Host:
Mouse