Immunocytochemical staining of mouse brain cortical neurons in culture. NFL immunoreactivity is RED and YELLOW. BLUE staining is DAPI nuclear staining. Anti-NFL (Aves Labs, 1:1000 dilution). Dr. Felix Eckenstein, University of Vermont.
Three antipeptide antibodies were generated in chickens against sequences shared between the mouse (P08551) and human (P07196) gene products. Antibodies were affinity-purified and mixed in equal concentrations. These antibodies have been validated with human, mouse and rat tissues.
Volume: 1000 µL
Concentration: 300 µg/mL
Host Species: Chicken
Species Reactivity: Human, Mouse, Rat
Applications: ICC, IHC, WB
Protein Name / Synonyms: Neurofilament light polypeptide (NF-L) (68 kDa neurofilament protein) (Neurofilament triplet L protein)
Target Description: Human Neurofilament Light Chain (NFL) is a 61,516 dalton protein (543 amino acids) expressed in the axons of CNS and PNS neurons, where it plays a role in maintaining the structural integrity of the axon. Mutations of the NFL gene product are responsible for some forms of autosomal dominant Charcot-Marie-Tooth neuropathies in humans (i.e., CMT type 1F).
Gene ID: NEFL NF68 NFL
Antibody Registry ID (RRID): AB_2313553
Physical State: Liquid
Production Notes: Chickens were immunized with three synthetic peptide / keyhole limpet hemocyanin (KLH) conjugates. These synthetic peptides corresponded to different regions of the NF-L gene product, but are shared between the human (P07196, NCBI) and mouse (P08551, NCBI) protein sequences. After repeated injections, immune eggs were collected, and the IgY fractions were purified from the yolks. These IgY fractions were then affinity-purified using a peptide column, and the concentrations of the eluates adjusted to 300 µg/mL. Finally, equal volumes of each of the three affinitypurified anti-peptide antibodies were mixed, and the preparation was filter-sterilized.
Validation and Application Notes
Molecular Weight: 70 kDa
Western Blot Dilution Range: 1:1000-1:2000
IHC Dilution Range: 1:1000-1:2000
Quality assurance analysis was performed using immunohistochemistry (at a dilution of 1:2000) using fluorescein-labeled goat anti-chicken IgY (1:500 dilution, Aves Labs Cat.# F-1005) as the secondary reagent.
Store at 4°C in the dark. Under these conditions, the antibodies should have a shelf life of at least 12 months (provided they remain sterile). Do not freeze these antibodies unless you want to store them for longer periods of time. Note, however, that each time an antibody preparation is frozen, about half of its binding activity is lost.
Aves Labs products are intended for use as research laboratory reagents. They are not intended for use as diagnostic or therapeutic reagents in humans.
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