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Key Documents

MAB353

Sigma-Aldrich

Anti-Nestin Antibody, clone rat-401

clone rat-401, Chemicon®, from mouse

Synonym(s):

rat401

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About This Item

UNSPSC Code:
12352203
eCl@ss:
32160702
NACRES:
NA.41

biological source

mouse

Quality Level

antibody form

purified antibody

antibody product type

primary antibodies

clone

rat-401, monoclonal

species reactivity

mouse, rat

should not react with

human

manufacturer/tradename

Chemicon®

technique(s)

immunocytochemistry: suitable
immunohistochemistry (formalin-fixed, paraffin-embedded sections): suitable
western blot: suitable

input

sample type neural stem cell(s)
sample type epithelial cells

isotype

IgG1

NCBI accession no.

UniProt accession no.

shipped in

wet ice

target post-translational modification

unmodified

Gene Information

human ... NES(10763)
mouse ... Nes(18008)
rat ... Nes(25491)

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

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

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

Target description

200-220 kDa

Physical form

Format: Purified
Protein A purified
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.

Analysis Note

Control
Mouse embryonic brain tissue

Other Notes

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

Legal Information

CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany

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.

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Storage Class

12 - Non Combustible Liquids

wgk_germany

WGK 1

flash_point_f

Not applicable

flash_point_c

Not applicable


Certificates of Analysis (COA)

Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.

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Designer self-assembling Peptide nanofiber scaffolds for adult mouse neural stem cell 3-dimensional cultures
Gelain, Fabrizio, et al.
Testing, 1, e119-e119 (2006)
Role of cyclin D1 cytoplasmic sequestration in the survival of postmitotic neurons.
Sumrejkanchanakij, P; Tamamori-Adachi, M; Matsunaga, Y; Eto, K; Ikeda, MA
Oncogene null
J Mokrý et al.
Folia biologica, 44(5), 155-161 (2000-03-25)
For a long time, intermediate filament protein nestin was recognized as a specific constituent of the cytoskeleton of developing neural cells and skeletal muscle. Recent reports by other investigators demonstrated that nestin may also be expressed in other cell types.
Markers of adult neural stem cells.
Eric Wexler
Methods in Molecular Biology null
Human embryonic stem cells express an immunogenic nonhuman sialic acid.
Martin, Maria J, et al.
Nature Medicine, 11, 228-232 (2005)

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Human iPSC neural differentiation media and protocols used to generate neural stem cells, neurons and glial cell types.

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