Nestin (ESTM46, FLJ21841, Intermediate Filament Protein, Nbla00170, Nes) (APC)

Antibodies / Antibodies-Neurofilament

4°C Do Not Freeze

Supplied as a liquid in PBS, pH 7.2. Labeled with Allophycocyanin (APC).

Nestin purified from embryonic rat spinal cord (P21263).

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): 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: 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:
Store product at 4°C in the dark. DO NOT FREEZE! Stable at 4°C for 12 months after receipt as an undiluted liquid. Dilute required amount only prior to immediate use. Further dilutions can be made in assay buffer. Caution: APC conjugates are sensitive to light. For maximum recovery of product, centrifuge the original vial prior to removing the cap.

Note: Applications are based on unconjugated antibody.