Recombinant Full Length Human NF-L

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