Name: | Recombinant Full Length Human NF-L |
HGNC Name: | NEFL |
RRID: | NA |
Format: | 1mg/mL in 6M urea, 10mM Phosphate pH=7.5 |
Applications: | Protein standard for ELISA, MSD, Luminex and Simoa assays, immunogen for antibody production |
Storage: | Stable at 4°C for several months. For longer term store at -20°C or lower |
Uniprot: | PO7196 |
Recombinant Human NF-L
Cat# PROT-r-NF-L
$300.00 – $2,000.00
Neurofilaments are the 10 nm or intermediate filament proteins found specifically in neurons, and are composed predominantly of four major proteins called NF-L, NF-M, NF-H and α-internexin. NF-L, NF-M and NF-H were named based on their apparent molecular weight on SDS-PAGE gels, so NF-L is low or light, NF-M is medium or middle and NF-H is high or heavy. On SDS-PAGE NF-L runs at 68-70kDa, NF-M at 145-160kDa and NF-H at 200-220kDa with some species variability, larger species tending to have larger molecules. In every case the real molecular weight is significantly lower since long acidic sequences in these molecules cause them to run aberrantly. These three proteins are major components of large diameter axons in the adult, while α-internexin is a more major component of the developing nervous system, although still present in the adult. NF-L and other neurofilament subunits accumulate in many neurological diseases, such as Lou Gehrig’s disease (ALS) and Alzheimer’s disease, and mutations in the protein coding region of the human NF-L gene cause some forms of Charcot-Marie-Tooth disease (2-4). NF-L is a very abundant protein particularly concentrated in large diameter axons and may leak into blood and CSF following various kinds of axonal injury and/or degeneration. There has therefore been much recent interest in the detection of NF-L in CSF and blood as a surrogate marker of neuronal damage and degeneration (5). NF-L is also known as NF-Light, Nfl and NEFL.
A codon optimized cDNA designed to express full length human neurofilament NF-L was inserted into pET30a(+) eukaryotic expression vector, which adds an N-terminal in frame His-tag and some other vector derived sequence. This was transformed into E. coli and the recombinant protein was purified in 6M urea using immobilized metal affinity chromatography. Purified protein was diluted to 0.5mg/mL and is supplied in 6M urea. Our protein preparation is widely used as a protein standard in ELISA, Simoa and other kinds of antibody based assays for NF-L detection. The product data and MSDS information can be downloaded from the “Data Sheets” tab below.
Human NF-L sequence was based on that was NP_006149.2 which was inserted into the eukaryotic expression vector pET30a(+) which adds an N terminal His-tag and some other sequence, underlined below. This sequence includes a thrombin cleavage site (blue), an S-tag affinity peptide (red) and an enterokinase cleavage site (green).
MHHHHHHSSG LVPRGSGMKE TAAAKFERQH MDSPDLGTDD DDKAMADIGS EFMSSFSYEP 60
YYSTSYKRRY VETPRVHISS VRSGYSTARS AYSSYSAPVS SSLSVRRSYS SSSGSLMPSL 120
ENLDLSQVAA ISNDLKSIRT QEKAQLQDLN DRFASFIERV HELEQQNKVL EAELLVLRQK 180
HSEPSRFRAL YEQEIRDLRL AAEDATNEKQ ALQGEREGLE ETLRNLQARY EEEVLSREDA 240
EGRLMEARKG ADEAALARAE LEKRIDSLMD EISFLKKVHE EEIAELQAQI QYAQISVEMD 300
VTKPDLSAAL KDIRAQYEKL AAKNMQNAEE WFKSRFTVLT ESAAKNTDAV RAAKDEVSES 360
RRLLKAKTLE IEACRGMNEA LEKQLQELED KQNADISAMQ DTINKLENEL RTTKSEMARY 420
LKEYQDLLNV KMALDIEIAA YRKLLEGEET RLSFTSVGSI TSGYSQSSQV FGRSAYGGLQ 480
TSSYLMSTRS FPSYYTSHVQ EEQIEVEETI EAAKAEEAKD EPPSEGEAEE EEKDKEEAEE 540
EEAAEEEEAA KEESEEAKEE EEGGEGEEGE ETKEAEEEEK KVEGAGEEQA AKKKD 595
Number of amino acids: 595
Molecular weight: 67224.80
Theoretical pI: 4.69
Amino acid composition:
Ala (A) 65 10.9%
Arg (R) 37 6.2%
Asn (N) 14 2.4%
Asp (D) 30 5.0%
Cys (C) 1 0.2%
Gln (Q) 29 4.9%
Glu (E) 102 17.1%
Gly (G) 25 4.2%
His (H) 12 2.0%
Ile (I) 20 3.4%
Leu (L) 50 8.4%
Lys (K) 41 6.9%
Met (M) 15 2.5%
Phe (F) 12 2.0%
Pro (P) 11 1.8%
Ser (S) 62 10.4%
Thr (T) 24 4.0%
Trp (W) 1 0.2%
Tyr (Y) 21 3.5%
Val (V) 23 3.9%
Total number of negatively charged residues (Asp + Glu): 132
Total number of positively charged residues (Arg + Lys): 78
Extinction coefficients are in units of M-1 cm-1, at 280 nm measured in water.
Ext. coefficient 36790
Abs 0.1% (=1 g/l) 0.547, assuming all pairs of Cys residues form cystines
Ext. coefficient 36790
Abs 0.1% (=1 g/l) 0.547, assuming all Cys residues are reduced
1. Hoffman et al. Neurofilament gene expression: a major determinant of axonal caliber. PNAS 84:3472-6 (1987).
2. Perrot R, et al. Review of the Multiple Aspects of Neurofilament Functions, and their Possible Contribution to Neurodegeneration.Mol. Neurobiol. 38:27-65 (2008).
3. Lépinoux-Chambaud C. Eyer J. Review on intermediate filaments of the nervous system and their pathological alterations. Histochem. Cell Biol. 140:13-22 (2013).
4. Liu Q. et al. Neurofilamentopathy in Neurodegenerative Diseases. Open Neurol. J. 5:58–62 (2011).
5. Bacioglu M, et al. Neurofilament light chain in blood and CSF as marker of disease progression in mouse models and in neurodegenerative diseases. Neuron 91:56-66 (2016).
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Contact info
EnCor Biotechnology Inc.
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Gainesville
Florida 32608 USA
Phone: (352) 372 7022
Fax: (352) 372 7066
E-mail: admin@encorbio.com