NESTIN; MSX-100UG

Code: MAB353 D2-231

Analysis Note

ControlMouse embryonic brain tissue

Application

Detect Nestin using this Anti-Nestin Antibody, clone rat-401 validated for use in I...


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Analysis Note

ControlMouse embryonic brain tissue

Application

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

Research CategoryStem Cell Research

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.

Research Sub CategoryNeural Stem Cells

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.

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.

Immunogen

Nestin purified from embryonic rat spinal cord.

Legal Information

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

Other Notes

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

Physical form

Protein A purified

Format: Purified

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

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

Specificity

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

Storage and Stability

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

Target description

200-220 kDa

antibody formpurified antibody
antibody product typeprimary antibodies
biological sourcemouse
clonerat-401, monoclonal
Gene Informationhuman ... NES(10763)mouse ... Nes(18008)rat ... Nes(25491)
inputsample type neural stem cell(s)sample type epithelial cells
isotypeIgG1
manufacturer/tradenameChemicon®
NCBI accession no.NP_037119.1
Quality Level100
shipped inwet ice
should not react withhuman
species reactivityrat, mouse
technique(s)immunocytochemistry: suitable, western blot: suitable, immunohistochemistry (formalin-fixed, paraffin-embedded sections): suitable
UniProt accession no.P21263
This product has met the following criteria: