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

Antibodies / Antibodies-Neurofilament

-20°C

Supplied as a liquid.

Raised against a segment of recombinant human Nestin expressed in bacteria and highly purified. The specific region was amino acids 317 to 630 of the human protein. This is a region which is 55% identical to rat Nestin and 57% identical to mouse Nestin.

Nestin is a member of the class IV intermediate filament protein family which is expressed in neuronal stem cells, which is the origin of the name nestin (1). Nestin was originally identified as a result of the production of a series of monoclonal antibodies directed against epitopes expressed on formalin fixed embyro day 15 rat spinal cord (2). One of these antibodies, called Rat 401, stained fibrous profiles in the developing nervous system, but not in the mature nervous system. By screening bacteriophage expression libraries with Rat 401, Lendahl et al. (1) were able to isolate a cDNA encoding the protein to which Rat 401 antibody bound. The protein proved to be an unusual member of the intermediate filament family, containing an alpha-helical region homologous to that found in keratin and neurofilament subunits. The protein was named Nestin as, in the developing nervous system, it is found in neural stem cells. The Human Genome Organization (HUGO) Gene Nomenclature Committee (HGNC) name for Nestin is NES. The Nestin protein has a very short N-terminal region in front of the alpha-helical region and a very long and repetitive C-terminal region. The molecular weight of human Nestin as determined by SDS-PAGE mobility is about 240kD. However the real molecular weight is considerably less than this, at 177kD, the disparity being likely due to the highly charged region of the C-terminal segment. Nestin is relatively poorly conserved in protein sequence across species boundaries, so that the mouse and human proteins have an overall identity of only 62%. As a result antibodies to the human protein often fail to recognize the rodent homolog and vice versa. However 031394 stains both rodent and human Nestin. Antibodies to Nestin are widely used to identify neural stem cells. The HGNC name for this protein is NES.

Applications:
Suitable for use in Immunocytochemistry and Western Blot. Other applications not tested.

Recommended Dilution:
Immunocytochemistry: 1:500
Western Blot: 1:5000; Expect to see a band at ~240kD and another much weaker at ~90kD, apparently a breakdown product of the ~240kD band.
Optimal dilutions to be determined by the researcher.

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 after receipt. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.