Western blot analysis of extracts from Neuro-2a, RD, and ZR-75-1 cells using Nestin Antibody (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).
|MW (kDa)||207-217 (human), 220-230 (mouse)|
Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA and 50% glycerol. Store at –20°C. Do not aliquot the antibody.
For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.
NOTE: Please refer to primary antibody product webpage for recommended antibody dilution.
From sample preparation to detection, the reagents you need for your Western Blot are now in one convenient kit: #12957 Western Blotting Application Solutions Kit
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
Load 20 µl onto SDS-PAGE gel (10 cm x 10 cm).
NOTE: Volumes are for 10 cm x 10 cm (100 cm2) of membrane; for different sized membranes, adjust volumes accordingly.
* Avoid repeated exposure to skin.
posted June 2005
revised June 2020
Protocol Id: 10
Nestin Antibody recognizes endogenous levels of total nestin protein.
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues near the carboxy terminus of human nestin protein. Antibodies are purified by peptide affinity chromatography.
Nestin is an intermediate filament family member protein that is structurally related to the neurofilament proteins (1). It is highly expressed in the developing brain, where it may help to regulate cell structure and intracellular processes required for neural cell division and migration (1,2). Upon maturation of the brain, nestin expression is quickly down-regulated and replaced by expression of the neurofilament proteins (2). Because nestin is expressed in both mature and precursor neuronal and glial cells, as well as in the developing brain and in the brain and spinal cord following damage, nestin is widely accepted as a marker of neural stem/progenitor cells (3). Research studies have shown that expression of nestin is also found in cells from various nervous system tumors, including gliomas, neuroblastomas, astrocytomas, and it is generally accepted as a marker for neural cancer stem cells (3). However, nestin expression has also been observed in astrocytes, retina, cardiac muscle, pancreas, and other tissues (3). Therefore, the acceptance of nestin as an exclusive marker of neural stem/progenitor cells is not unanimous.
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