Anti-NF-L Antibody [1B11] (A104310)

Immunostaining of a coronal section of the spinal cord of a rat given a midline C4 contusion injury three days previously. Sections were stained with Anti-NF-L Antibody (A270580) (red) and Anti-NF-L Antibody [1B11] (A104310) (green). Anti-NF-L Antibody [1B11] (A104310) stains prominent aggregates of material concentrated in the lateral funiculi and the dorsal columns but seen in lesser amounts throughout the section. These are degenerating and degenerated axons damaged by the C4 lesion. The Anti-NF-L Antibody antibody binds the C-terminal “tail” region of NF-L which is absent or destroyed during degeneration, so the positive profiles are largely negative for Anti-NF-L Antibody.

Immunohistochemistry analysis of a formalin fixed paraffin embedded brain stem section from a transgenic mouse model of ALS stained with Anti-NF-L Antibody [1B11] (A104310) at a dilution of 1:1,000 detected in DAB (brown) following the Vector Labs Mouse on Mouse ImmPRESS® HRP method, without the antigen retrieval step. Counterstained with Hematoxylin (blue). The Anti-NF-L Antibody [1B11] (A104310) labels what are clearly degenerated axons, showing typical swollen, sinusoidal and discontinuous profiles. Note that under these conditions healthy axons are not stained.

Western blots of Uman NF-LIGHT™ antibodies and a set of reagents on Recombinant Human NF-L Protein (A270573) and Recombinant Human NF L Protein (A333288). The lanes contain: [Lane 1] protein standards (red), [Lane 2] Recombinant Human NF-L Protein (A270573), [Lane 3] Recombinant Human NF L Protein (A333288). Recombinant Human NF-L Protein (A270573) runs at about 75kDa, while Recombinant Human NF L Protein (A333288) runs at about 12kDa. All five antibodies recognize both constructs. UD1 is also known as 2.1 is the detection reagent in the Uman NF-LIGHT™ assay while UD2, also known as 47.3 is the capture reagent (6). The three other lanes show results obtained with Anti-NF-L Antibody [1B11] (A104310) and Anti-NF-L Antibody [6H63] (A333276) respectively as indicated. All these antibodies binds to an epitope flanking the so-called second “stutter” in the Coil 2 region of the a-helical coiled coil “rod” region of NF-L. The binding properties are very similar to the Uman mouse monoclonal antibody UD1 also known as 2.1, as described originally in Norgren et al. 2002. This antibody is the key capture reagent in the NF-Light™ assay of Uman Diagnostics and the Quanterix Simoa™ NF-L assay.