Neurofilament M (Neurofilament Medium Polypeptide, NF-M, Neurofilament Triplet M Protein, 160kD Neurofilament Protein, Neurofilament 3, Nefm, Nef3, Nfm)

Pab

IgY

-20°C

Supplied as a liquid, 0.02% sodium azide.

C-terminal extension of rat NF-M, the so-called KE segment, was expressed in bacteria and purified from inclusion bodies by dissolving in 6M urea followed by preparative gel electrophoresis.

Neurofilaments can be defined as the intermediate or 10nm filaments found in specifically in neuronal cells. In the electron microscope neurofilaments appears as 10nm diameter fibres of indeterminate length which generally have fine wispy protrusions from their sides. They are found particularly abundantly in axons of large projection neurons. They probably function to provide structural support for neurons and their synapses and to support the large axon diameters required for rapid conduction of impulses down axons. They are composed a mixture of subunits which usually include the three neurofilament triplet proteins, known as NF-L, NF-M and NF-H. Neurofilaments may also include smaller amounts of peripherin, a-internexin, nestin and in some cases vimentin. Antibodies to the various neurofilament subunits are very useful cell type markers since the proteins are among the most abundant of the nervous system, are expressed only in neurons, and are biochemically very stable. To raise this particular antibody the C-terminal extension of rat NF-M, the so-called KE segment, was expressed in bacteria and purified from inclusion bodies by dissolving in 6M urea followed by preparative gel electrophoresis (1). This antibody was generated in chicken by standard procedures and immunoglobulin was extracted from egg yolk. The resulting polyclonal antibody belongs to the IgY subclass. This is the chicken homologue of mammalian IgG and can be used in the same general way, with the caveat that this type of antibody does not bind either Protein A or Protein G. The HGNC name for this protein is NEFM.

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

Recommended Dilutions:
Immunofluorescence (IC): 1:2000
Immunohistochemistry: 1:2000
Western Blot: 1:2000-1:5000
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.