Anti-Nestin Antibody, clone rat-401

200-220 kDa

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.

Nestin purified from embryonic rat spinal cord.

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.

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

Format: Purified

Protein A purified

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

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

Control
Mouse embryonic brain tissue

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

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

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