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Mouse monoclonal antibody to Neurofilament Light [DA2]


Catalogue No. M-1391-50
Description Neurofilaments are composed of three intermediate filament proteins: light (~68 kDa), medium (~160 kDa) and heavy (~200 kDa), which are involved in the maintenance of the neuronal caliber. Neurofilament light (NF68 or NF-L) is the most abundant of the three proteins.
Batch No. See product label
Unit size 50 uL
Antigen A preparation of enzymatically dephosphorylated pig neurofilaments including NF-L, NF-M and NF-H. Screening was by ELISA on the immungen followed by immunofluorescence microscopy. The epitope for this antibody is not in the N-terminus but its exact location is not known. It is thought to be somewhere within the central alpha helical "rod" region of the molecule.
Antibody Type Monoclonal
Isotype IgG1
Clone DA2
Other Names NF-L; NF68; NEFL; Neurofilament light polypeptide; NFL;
Accession P02547 NFL_PIG; P07196 NFL_HUMAN;
Produced in Mouse
Applications Western Blotting (WB), Immunocytochemistry (IC), Immunohistochemistry (IH) and Flow Cytometry (2 ug per 10^6 cells). A dilution of 1:5,000 - 1:10,000 is recommended for WB. A dilution of 1:100 - 1:500 is recommended for IC and IH. Biosensis recommends optimal dilutions/concentrations should be determined by the end user.
Specificity Specifically recognizes the light neurofilament subunit NF-L (~68 kDa) in WB.
Antibody Against Neurofilament Light
Cross-reactivity Hu, Rat, Ms, Fel, Bov, Por, Chk
Blast URL Click here
Form Lyophilised with 5% trehalose
Appearance White powder
Reconstitution Reconstitute in sterile distilled water. Centrifuge to remove any insoluble material.
Storage After reconstitution of lyophilised antibody, aliquot and store at -20C for a higher stability. Avoid freeze-thaw cycles.
Expiry Date 12 months after purchase
Specific References 1. Felitsyn N. et al (2008) The heme precursor delta-aminolevulinate blocks peripheral myelin formation. J Neurochem. 2008 Sep;106(5):2068-79.
Images (click to zoom)
Mouse monoclonal antibody to Neurofilament Light [DA2] Left: Cells grown from adult rat brain. The large cell in the middle is stained with Mouse monoclonal antibody to Neurofilament Light [DA2] M-1391-50 (green). Another type of neuronal lineage cell was stained with Rabbit polyclonal to alpha-internexin R-1379-50 (red). These cells were mitotic but had several characteristics of neurons. Right: Analysis of neurofilament, light, 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 ug/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.
Mouse monoclonal antibody to Neurofilament Light [DA2] Left: Visualization of Neurofilament Light expression in rat frontal cortex by Immunohistochemistry. Section was stained with mouse anti-NF-L antibody (red, 1:500), and co-stained with chicken antibody to GFAP (C-1373-50, 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 sections were 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: Western blot analysis of NF-L expression in whole tissue lysates (green, 1:5,000) in green: [1] protein standard, [2] rat brain, [3] rat spinal cord, [4] mouse brain, [5] mouse spinal cord. One strong band at 68-70 kDa corresponds to the NF-L protein.
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