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.

Reacts with human Nestin, with weak reactivity to monkey nestin. Not reactive with rodent nestin.

Epitope: human

Fusion protein.

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.

Research Category
Stem Cell Research

Research Sub Category
Neural Stem Cells

This Anti-Nestin Antibody, clone 10C2 is validated for use in IC, IH, IH(P), WB for the detection of Nestin.

200-220 kDa

Replaces: AB5922

Format: Purified

Protein A Purified mouse immunoglobulin in 20 mM sodium phosphate, 250 mM NaCl, pH. 7.6, with 0.1% sodium azide as a preservative.

Protein A purified

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.

Control
Positive Control: WI-38 (ATCC# CCL75) or U251 cell lines

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

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