Neurofilament light polypeptide (NF-L), Mouse Monoclonal Antibody (DA2)

Image of a neuron in cell culture, derived from adult rat cortex, by Immunofluorescence and Immunohistochemistry. The large middle cell is stained with M-1391-50, Neurofilament light polypeptide (NF-L), Clone DA2, Mouse mAb, (green), and co-stained with product R-1379-50, Alpha-internexin, Rabbit pAb, (red). The culture was grown under conditions to induce neuronal survival and differentiation, and product R-1379-50 highlights a network of small neurons at early stages of differentiation.

Analysis by western blot of NF-L expression in different tissue lysates with M-1391-50, Neurofilament light polypeptide (NF-L), Clone DA2, Mouse mAb, (green, 1:5,000) with strong banding at 68-70kDa corresponding to the NF-L protein. Lane 1: protein standard (red); Lane 2: rat brain; Lane 3: rat spinal cord; Lane 4: mouse brain; Lane 5: mouse spinal cord.

Left: Image of a rat frontal cortex section by Immunofluorescence and Immunohistochemistry. The section was stained with M-1391-50, Neurofilament light polypeptide (NF-L), Clone DA2, Mouse mAb, (red, 1:500), and co-stained with product C-1373-50, Glial fibrillary acidic protein (GFAP), Chicken pAb, (green, 1:5,000). IHC Method: Following transcardial perfusion of rat with 4% paraformaldehyde, brain was post-fixed for 24 hours, cut into 45 um sections, and free-floating section was stained. The NF-L antibody labels cell bodies and processes of pyramidal neurons, as well as dendrites and axons of other neuronal cells, while the GFAP antibody stains the network of glial cells. Right: Analysis of Neurofilament light polypeptide (NF-L), expression in human prostate cancer cell line DU145 by flow cytometry. Fixing and Permeabilization of cells: Absolute methanol (10 minutes in ice) and 0.1% Tween-20 in PBS, Blocking: 200 µg/mL Normal Sheep IgG (20 minutes), Primary antibody: Mouse Monoclonal antibody to Neurofilament Light (cat # M-1391-50, 2 μg per ~10^6 cells) for 30 minutes at room temperature, Secondary antibody: Goat anti-mouse PE labeled secondary antibody (1:100 fold dilution) with incubation for 20 minutes in dark at room temperature. Non-specific Control IgG, clone X63 (cat # M-1249-200) was used as negative control under same conditions (black dashed). Flow cytometry data and results were generated using Orflo MoxiflowTM instrument and protocols.