Rat Neurofilament light polypeptide/NEFL ELISA Kit

From Assay Genie, this Sandwich ELISA, Double Antibody kit is designed for the accurate quantification of Neurofilament light polypeptide (NEFL) levels in rat serum, plasma, cell culture supernatant, cell or tissue lysate, and other liquid samples. NEFL is a cytoskeleton protein found in neurons of the peripheral and central nervous system, where it plays a role in determining axonal calibre and supporting axonal structure and function. Detection is colorimetric, with a sensitivity of 9.375 pg/mL and a working range of 15.625–1000 pg/mL. Synonyms for the target include NEFL, NF-L, 68 kDa neurofilament protein, and Neurofilament triplet L protein (UniProt P19527). Kit components should be stored at 2–8°C, where the kit is stable for 12 months.

Changes in NEFL levels have been associated with several neurological disorders, including Alzheimer's disease, multiple sclerosis, traumatic brain injury, Amyotrophic Lateral Sclerosis, Parkinson's disease, and Charcot-Marie Tooth disease, making it a valuable biomarker for neurodegenerative research. The assay procedure involves equilibrating reagents and TMB substrate to room temperature, loading standards, samples and blanks onto the pre-coated plate, primary incubation at 37°C for 90 minutes, biotin-labelled detection antibody incubation at 37°C for 60 minutes, HRP-Streptavidin binding at 37°C for 30 minutes, TMB colour development in the dark for 10–20 minutes, and a stop reaction with absorbance reading at 450 nm.

Sample preparation guidance covers serum (clot and centrifuge at 1000 × g for 20 minutes), plasma (anticoagulant tubes, centrifuge at 1000 × g for 15 minutes at 2–8°C), tissue homogenate (homogenise in PBS plus protease inhibitors), cell culture supernatant (centrifuge at 2500 rpm for 5 minutes) and cell lysates (lysis buffer with protease inhibitors). Reported linearity for serum, EDTA plasma and heparin plasma (n = 5) at 1:2, 1:4 and 1:8 dilutions falls between 81% and 104%. Recovery across serum, EDTA plasma and heparin plasma ranges from 85% to 105%, with averages between 90% and 95%.