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  • MAB353 - Anti-Nestin Antibody, clone rat-401

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MAB353 Sigma-Aldrich

Anti-Nestin Antibody, clone rat-401

Detect Nestin using this Anti-Nestin Antibody, clone rat-401 validated for use in IC, IH, IH(P) & WB.

Synonym: rat401

  •  eCl@ss 32160702

  •  NACRES NA.41

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Properties

Related Categories Alphabetical Index, Antibodies, NE-NI, Primary Antibodies
clone   rat-401, monoclonal
biological source   mouse
application(s)   immunocytochemistry: suitable
  immunohistochemistry: suitable (paraffin)
  immunohistochemistry: suitable
  western blot: suitable
species reactivity   rat, mouse
should not react with   human
shipped in   wet ice
isotype   IgG1
antibody product type   primary antibodies
Quality Level   100
mfr. no.   Chemicon®
input   sample type neural stem cell(s)
sample type epithelial cells
NCBI accession no.   NP_037119.1
UniProt accession no.   P21263

Description

General description

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 (Mokry and Nemecek, 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 carcinomas.

Specificity

Rodent Nestin protein confirmed with rat and mouse. Rat-401 is not reactive with human nestin protein.

Immunogen

Nestin purified from embryonic rat spinal cord.

Application

Research Category
Stem Cell Research

Research Sub Category
Neural Stem Cells

Western Blot:
200-220 kDa in reducing gels of newborn rat or mouse cell extracts. Note that nestin was originally reported (erroneously) to migrate at approximately 440 kDa by SDS-PAGE (Kachinsky, 1995).
For Westerns, prepare samples by boiling in 4 volumes of 125 mM Tris, pH 6.8, 10% 2-mercaptoethanol, 10% glycerol and 4.6% SDS. 5% PAGE gels are suggested. Block membranes with milk or BSA. Do not dilute primary antibody in buffer containing milk. Freezing and thawing of samples or samples not stored at -85°C will show breakdown of nestin sometimes into multiple fragments.

Immunohistochemistry:
1:20-1:200 dilution of a previous lot was used. Perfuse tissue with 4% paraformaldehyde at pH 7.4 for light microscopy or with either 4% paraformaldehyde at pH 10.0 or 4% paraformaldehyde with 0.1% glutaraldehyde at pH 7.4 for EM. Suitable for frozen or paraffin-embedded sections.

Immunocytochemistry:
1:20-1:200 dilution of a previous lot was used. Fix cells in 4% paraformaldehyde buffered with 50 mM sodium borate at pH 9.5.

Optimal working dilutions and protocols must be determined by end user.

Target description

200-220 kDa

Physical form

Format: Purified

Protein A Purfied

Purified mouse monoclonal IgG1 in buffer containing 0.09% sodium azide.

Storage and Stability

Stable for 1 year at 2-8ºC from date of receipt.

Disclaimer

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.

Quality

Immunohistochemistry(paraffin):
Nestin (cat. # MAB353, Clone 401) staining of Mouse Pancreas, tissue was pretreated with citrate buffer, pH 6.0. A previous lot of this antibody was diluted to 1.25 µg/mL, using IHC-Select detection with HRP-DAB. Immunoreactivity is detected in the islets of Langerhans.
Optimal Staining of Nestin Monoclonal Antibody: Mouse Pancreas

Analysis Note

Control
Mouse embryonic brain tissue

Other Notes

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

Safety & Documentation

Safety Information

Safety Information for this product is unavailable at this time.
Protocols & Articles

Articles

Antibody Basics

Immunoglobulins (Igs) are produced by B lymphocytes and secreted into plasma. The Ig molecule in monomeric form is a glycoprotein with a molecular weight of approximately 150 kDa that is shaped more ...
Keywords: Affinity chromatography, Centrifugation, Chromatography, Digestions, Direct immunofluorescence, Gene expression, High performance liquid chromatography, Immunofluorescence, Ion Exchange, Microscopy, Precipitation, Purification, Rheumatology, Scanning electron microscopy

Protocols

Neural Stem Cell Culture Protocols

Introduction Methods     Isolation of Neural Stem Cells         Isolation of NSCs from Neural Tissue         Differentiation of Human iPSCs into NSCs         Characterization of Neural Stem Cells    ...
Keywords: AGE, Adhesion, Apoptosis, Cell culture, Central Nervous System, Cryopreservation, Culture media, Diseases, Growth factors, High performance liquid chromatography, Phase transitions, Phosphorylations, Purification

Western Blot Protocol | Immunoblotting Protocol

Western Blotting refers to the electrophoretic transfer of proteins from sodium dodecyl sulfate polyacrylamide gels to sheets of PVDF or nitrocellullose membrane, followed by immunodetection of prote...
Keywords: AGE, Buffers, Cell disruption, Detection methods, Detergents, Dialysis, Electroblotting, Electrophoresis, Enzyme activity, Gel electrophoresis, Immunoprecipitation, PAGE, Protein extraction, Purification, Sample preparations, Western blot

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Peer-Reviewed Papers
15

References

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