Technical Data
N2060-10
Nestin (ESTM46, FLJ21841, Intermediate Filament Protein, Nbla00170, Nes)
Description:
Nestin, a large intermediate filament protein (class Type VI), is 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 primative 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.

Applications:
Suitable for use in Immunohistochemistry, Western Blot and Immunocytochemistry. Other applications have not been tested.

Recommended Dilutions:
Immunohistochemistry (Frozen or paraffin): 1:20-1:200. Perfuse tissue with 4% paraformaldehyde, pH 7.4 for light microscopy or with either 4% paraformaldehyde, pH 10 or 4% paraformaldehyde with 0.1% glutaraldehyde, pH 7.4 for EM.
Western Blot: 200-220kD in reducing gels of newborn rat or mouse cell extracts. Prepare samples by boiling in 4 volumes of 125mM Tris, pH 6.8, 10% 2-mercaptoethanol, 10% glycerol, 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 -70°C will show breakdown of nestin sometimes into multiple fragments.
Immunocytochemistry: 1:20-1:200. Fix cells in 4% paraformaldehyde buffered with 50mM sodium borate, pH 9.5.
Optimal dilutions to be determined by the researcher.

Recommended Positive Control:
Mouse embryonic brain tissue

Storage and Stability:
May be stored at 4°C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20°C. Aliquots are stable for 12 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
TypeIsotypeCloneGrade
MabIgG12Q178Purified
SizeStorageShippingSourceHost
100ug-20°CBlue IceRatMouse
Concentration:
~1mg/ml
Immunogen:
Nestin purified from embryonic rat spinal cord.
Purity:
Purified
Form
Supplied as a liquid in Tris-Glycine, 0.15M sodium chloride, pH 7.4, 0.05% sodium azide.
Specificity:
Recognizes rodent Nestin at 200-220kD. Species Crossreactivity: mouse and rat. Does not crossreact with human.
Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.
1. Bossolasco, P., et al., Experimental Neurology 193: 312-325 (2005). 2. De Hemptinne, I., et al., J. Neurochemistry 91: 155-166 (2004). 3. Niu, S., et al., Neuron 41: 71-84 (2004). 4. Noctor, S., et al., Nat. Neurosci. 7: 134-144 (2004). 5. Harry, G., et al., Neurotox. Res. 5: 623-627 (2004). 6. Scemes, E., et al., J. Neurosci. 23: 11,444-11,452 (2003). 7. Ehtesham, M., et al., Cancer Research 62: 5657-5663 (2002). 8. Safford, K.M., et al., Biochem. Biophys. Res. Comm. 294(2): 371-379 (2002). 9. Duittoz, A.H. & Hevor, T., J. Neuroscience Methods 107: 131-140 (2001). 10. Mokry, J. & Nemecek, S., Folia Biologica 44: 155-161 (1998). 11. Gritti, A., et al., J. Neurosci. 16: 1091-1100 (1996). 12. Kachinsky, A., et al., J. Histochem. Cytochem. 43(8): 843-847 (1995). 13. Rosen G., et al., Dev. Brain Res. 82: 127-135 (1994). Okada,S., et al Nature Medicine (2006) 12: 829-834.