Name: Recombinant Human Neurofilament light polypeptide (NEFL)
Brand: CUSABIO
SKU: CSB-EP015688HU

Product Details

Purity

Greater than 90% as determined by SDS-PAGE.

Target Names

NEFL

Uniprot No.

P07196

Research Area

others

Alternative Names

NEFL; NF68; NFL; Neurofilament light polypeptide; NF-L; 68 kDa neurofilament protein; Neurofilament triplet L protein

Species

Homo sapiens (Human)

Source

E.coli

Expression Region

2-543aa

Target Protein Sequence

SSFSYEPYYSTSYKRRYVETPRVHISSVRSGYSTARSAYSSYSAPVSSSLSVRRSYSSSSGSLMPSLENLDLSQVAAISNDLKSIRTQEKAQLQDLNDRFASFIERVHELEQQNKVLEAELLVLRQKHSEPSRFRALYEQEIRDLRLAAEDATNEKQALQGEREGLEETLRNLQARYEEEVLSREDAEGRLMEARKGADEAALARAELEKRIDSLMDEISFLKKVHEEEIAELQAQIQYAQISVEMDVTKPDLSAALKDIRAQYEKLAAKNMQNAEEWFKSRFTVLTESAAKNTDAVRAAKDEVSESRRLLKAKTLEIEACRGMNEALEKQLQELEDKQNADISAMQDTINKLENELRTTKSEMARYLKEYQDLLNVKMALDIEIAAYRKLLEGEETRLSFTSVGSITSGYSQSSQVFGRSAYGGLQTSSYLMSTRSFPSYYTSHVQEEQIEVEETIEAAKAEEAKDEPPSEGEAEEEEKDKEEAEEEEAAEEEEAAKEESEEAKEEEEGGEGEEGEETKEAEEEEKKVEGAGEEQAAKKKD
Note: The complete sequence including tag sequence, target protein sequence and linker sequence could be provided upon request.

Mol. Weight

81.4kDa

Protein Length

Full Length of Mature Protein

Tag Info

N-terminal 10xHis-SUMO-tagged and C-terminal Myc-tagged

Form

Liquid or Lyophilized powder
Note: We will preferentially ship the format that we have in stock, however, if you have any special requirement for the format, please remark your requirement when placing the order, we will prepare according to your demand.

Buffer

If the delivery form is liquid, the default storage buffer is Tris/PBS-based buffer, 5%-50% glycerol.
Note: If you have any special requirement for the glycerol content, please remark when you place the order.
If the delivery form is lyophilized powder, the buffer before lyophilization is Tris/PBS-based buffer, 6% Trehalose.

Reconstitution

We recommend that this vial be briefly centrifuged prior to opening to bring the contents to the bottom. Please reconstitute protein in deionized sterile water to a concentration of 0.1-1.0 mg/mL.We recommend to add 5-50% of glycerol (final concentration) and aliquot for long-term storage at -20°C/-80°C. Our default final concentration of glycerol is 50%. Customers could use it as reference.

Troubleshooting and FAQs

Protein FAQs

Storage Condition

Store at -20°C/-80°C upon receipt, aliquoting is necessary for mutiple use. Avoid repeated freeze-thaw cycles.

Shelf Life

The shelf life is related to many factors, storage state, buffer ingredients, storage temperature and the stability of the protein itself.
Generally, the shelf life of liquid form is 6 months at -20°C/-80°C. The shelf life of lyophilized form is 12 months at -20°C/-80°C.

Lead Time

3-7 business days

Notes

Repeated freezing and thawing is not recommended. Store working aliquots at 4°C for up to one week.

Datasheet & COA

Please contact us to get it.

Description

The recombinant human neurofilament light polypeptide (NEFL) is prepared in the E.coli. Its expression region lies in the fragment encoding amino acid residues 2-543 of the human NEFL protein. This recombinant human NEFL protein is labeled with a 10xHis-SUMO-tag at the N-terminus and a Myc-tag at the C-terminus. It harbors the full length of the mature protein in length. The purity of this NEFL protein is greater than 90% determined by SDS-PAGE. On the gel, this protein migrated to the band with a molecular weight of about 90 kDa. And its component has been identified by the LC-MS/MS Analysis.

The NEFL protein is the light chain of neurofilaments that play an important role in axonal growth and the determination of axonal caliber. As axons are injured and die in neurodegenerative processes, NEFL leaks into the interstitial space, then into CSF and plasma. NEFL levels thus increase as neurodegenerative diseases progress and can reflect disease activity. Numerous studies have shown that NEFL is a potential biomarker for determining the stage of disease, tracking progression, and aiding in the identification of disease-modifying treatments in neurological disorders.

Customer Reviews and Q&A

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■ Q&A

Concerning the Recombinant Human Neurofilament light polypeptide, I have a question about the tag(s) and the possibility to obtain the protein with a free N-terminus ( if possible also C-terminal free). From the SUMO-tag, There exist the possibility to remove by protease at the end of the purification process. Can you check with Cusabio’s R&D people which possibilities exist?

The protein of CSB-EP015688HU that we currently developed is N-terminal 10xHis-SUMO-tagged and C-terminal Myc-tagged.
If you need to remove the tag or tag-free protein, please communicate with us in advance. For the C-terminal tag, we can definitely guarantee to remove, however, for the N-terminal tag, we can try enzyme digestion, but we can't guarantee 100% successfully. . Not all protein tags can be removed as some proteins will be very unstable after tag removal.
If we succeed in removing the tag, we will charge an extra cost.
If we fail in removing the tag, we won’t charge for any extra cost and remark this information in the datasheet as follows “Note: The laboratory determined that the Tag on your protein could not be removed with standard laboratory procedures. Your protein is being supplied with the Tag intact.”
Generally, the delivery time will be extended for 3 days.

We order this protein CSB-EP015688HU several times and are satisfied with the test result, is there any discount for bulk orders?

Yes, we can offer an additional discount for bulk orders, please let us know your demanded quantity and your requirement for the package and shipping format.

I'm experiencing difficulties with measuring protein using nanodrop A280. I was wondering if you have tested the protein concentration of the Recombinant 68kDa filament protein (CSB-EP015688HU) using this technique?

We use the Bradford method to detect its concentration. Glycerol has a strong interference to the NanoDrop 2000c detection, even with a final concentration of 10% glycerol (you can refer to this report: http://www.sislab.info/NPhP_compa_ND.pdf ). A280 can be measured before adding glycerol. After adding glycerol, use the measured value before doing a conversion. If the protein you ordered is in liquid form, it should have some glycerol added, so it will encounter difficulties. It is recommended that you can try another concentration detection method to test again, such as the Bradford method.

Could you please help to check if isoelectric point information of product CSB-EP015688HU is available?

CSB-EP015688HU Theoretical pI: 4.89

Does CSB-EP015688HU contain imidazol?

We have performed dialysis to change the buffer, and the imidazole is removed, but the final situation is not tested, so there is no guarantee that there will be no imidazole at all.

Is it possible to provide a small size as 5ug for CSB-EP015688HU? And do you also provide this protein from Mouse or Rat species?

Yes, we could offer a small size of 5ug as a free sample for you to test as long as you can share the test report with us, please provide your shipping information and application for this protein.
Yes, we can offer this protein from Mouse and Rat species. Please go to the following product pages to check them.

https://www.cusabio.com/Recombinant-Protein/Recombinant-Mouse-Neurofilament-light-polypeptideNefl-Yeast-19605.html
Recombinant Mouse Neurofilament light polypeptide(Nefl)
CSB-YP015688MO >> Yeast
CSB-EP015688MO >> E.coli
CSB-BP015688MO >> Baculovirus
CSB-MP015688MO >> Mammalian cell

https://www.cusabio.com/Recombinant-Protein/Recombinant-Rat-Neurofilament-light-polypeptide-Nefl--5739.html
Recombinant Rat Neurofilament light polypeptide(Nefl)
CSB-YP015688RA >> Yeast
CSB-EP015688RA >> E.coli
CSB-BP015688RA >> Baculovirus
CSB-MP015688RA >> Mammalian cell

Are both tags, i.e. 10xHis-SUMO tag at the N-terminus and Myc tag at the C-terminus used for purification? Is the SUMO tag removed by SUMO Protease treatment?

Thanks for your inquiry. Here is the informaton about tags:
①　The His changes from 6*his to 10*his. The main reason is that 10*His has a stronger affinity for the column material whih is helpful for those proteins that can't attach to the column very well.
It is also helpful for protein purification (The specific adsorption capacity target protein is strong, and the nonspecific adsorption ability of the impurity protein is weak, then we can design a high concentration of midazolam to further remove the impurity protein).
②　We add myc-tag at C-terminus. This is mainly for the convenience of customers and we to do WB detection for myc-tag antibody, thus further test the accuracy of the target protein. Although we can also detect His-tag antibodies by WB, but relatively speaking, its specificity is not good as myc-tag.
③　We add thrombin site for all tags. It will be more convenient if you require tag removal later.

I was wondering if you could let me know at what concentration the current lot of CSB-EP015688HU / Recombinant Human Neurofilament light polypeptide(100 µg) is provided.

Thanks for your inquiry. Regarding this item, CSB-EP015688HU, we have the protein both in liquid form and lyophilized form currently.
Liquid form: 0.32mg in stock. Lot number is 03193. The concentration is 0.8mg/ml.
Lyophized form: 2.44mg in stock. Lot number is 03403. The concentration before lyophization is 0.6mg/ml. You could redissolve it to your wanted concentration.

Has your protein CSB-EP015688HU already passed the ELISA test?

The proteins that we currently have in stock all passed the ELISA verification.

We are looking for a NEFL Recombinant protein (Human) that is suitable to use as a standard for an ELISA. Is the CSB-EP015688 suitable? If yes, would be able to recommend an antibody pair that has been optimized in ELISA for this protein?

There are many ELISA experimental factors, which are related to your experimental platform and technology, and each experimental stage may affect the experimental results. It is recommended that you use the corresponding expression system to express protein and antibody detection, and first purchase small specifications for testing. Sorry, at the same time, we have no corresponding Antibody pair recommendation.

Can you also confirm that these sources are carrier protein free?

All of our proteins are carrier-free.

For this protein CSB-EP015688HU, is there any affinity tag on the protein?

Yes, there is an affinity tag (His-sumo tag) on the protein.

Is this protein soluble?

We offer five expression systems: E. coli, Yeast, Baculovirus, Mammalian cell, and Cell-Free (in vitro E.coli). Among them, yeast only has supernatant expression, and the protein expressed in the supernatant must be a soluble protein.
The E. coli, Baculovirus, Mammalian cell, and Cell-Free (in vitro E.coli) have both supernatant expression and inclusion body expression, as long as the protein expressed in the supernatant, it must be soluble, if it is the protein expressed by in the inclusion body, we will also take a variety of methods to refold it to finally ensure that all the proteins we provide are soluble.
So all the proteins we offer are soluble.

How to reconstitute this protein CSB-EP015688HU?

Please use deionized sterile water to reconstitute proteins. (Dissolve completely. The concentration could depend on your experiment demand.)
We suggest that the concentration is not lower than 0.1mg/ml. Proteins will be unstable if they're too diluted. Generally, we recommend 0.1-1.0mg/ml concentration. If you have special requirements for the concentration, you could dilute the protein according to their experiment demand.
We haven’t studied whether the optimal concentration range will vary among different hosts such as e.coli, yeast, mammalian, or baculovirus, so we can’t provide usable advice.
If you need to use it immediately after dissolving one protein, you don't need to add glycerol. If you won't use the protein in short term, we suggest adding some glycerol for storage. Reconstituting a suitable amount of protein and store the remaining protein in lyophilized powder is recommended. You could remark "The lyophilized powder should be packed into multiple vials" when placing orders. The amount of lyophilized powder in each vial will depend on your experiment demand.
In principle, the final concentration of glycerol from 20% to 50% will be OK. Our default final concentration of glycerol is 50%. You could use it as a reference.

Have you validated the activity of this protein? If not, how to express a protein with bioactivity?

This protein is not included in our activity verification plan currently, we haven’t tested the activity of this protein yet, you need to empirically determine if it has protein activity.
A protein from the expression to the establishment of the activity test system to complete the activity verification requires a lot of cost and time investment. We are sorry that we have not tested it and do not guarantee activity. In theory, as what we provide is the full length of the mature protein, and we adopt affinity chromatography for purification, which is purified under a very mild condition, it's very likely to be active. We recommend that you can order a small size to have a try.
For gaining a protein with bioactivity, you should choose a right expression system, a suitable expression vector, an appropriate purification method, and a validation experiment. You can learn more from this link: https://www.cusabio.com/c-20275.html.
We will refer to the relevant literature on the preparation method of the active protein on the market, or if you have consulted the relevant literature and hope that we can prepare according to the methods in the literature, we will ensure the activity to the maximum according to the preparation methods mentioned in the literature.

Target Background

Function

Neurofilaments usually contain three intermediate filament proteins: NEFL, NEFM, and NEFH which are involved in the maintenance of neuronal caliber. May additionally cooperate with the neuronal intermediate filament proteins PRPH and INA to form neuronal filamentous networks.

Gene References into Functions

NF-L may be involved in severity of neuronal injury following traumatic brain injury PMID: 27819296
CSF neurofilament light chain protein is an accurate marker for distinguishing Alzheimer's disease patients from healthy controls. PMID: 28527630
Cerebrospinal fluid neurofilament protein light is a useful tool for determining disease intensity in frontotemporal dementia and motor neuron disease patients. PMID: 28631955
Our results suggest that plasma NFL levels may not be a useful biomarker for the diagnosis of prodromal and dementia stages of AD. PMID: 28428015
In this Chinese Han population a novel Charcot-Marie-Tooth disease-associated gene mutations of NEFL (c.280C>T) was discovered. PMID: 27862672
We conclude that low BDNF and high LCN2 and NF-L levels are associated with Multiple Sclerosis (MS) pathogenesis, and high IGFBP1level is a biomarker for female MS only, suggesting different MS progression pathways between the sexes. LCN2 is a candidate predictor of response to natalizumab treatment, and NF-L is a candidate predictor of clinically isolated syndrome's (CIS) conversion into MS. PMID: 29108879
This study showed tat Serum NfL concentration is increased in familial Alzheimer disease prior to symptom onset and correlates with measures of disease stage and severity. Serum NfL may thus be a feasible biomarker of early AD-related neurodegeneration. PMID: 29070659
Results show that both alphaS and NFL can be phosphorylated by CKII, PLK2 and PLK3, but Ser129 in alphaS is a preferential site for PLK2 and PLK3, demonstrating higher phosphorylation efficiency. Comparatively, CKII preferentially phosphorylates Ser473 in NFL and this site can be phosphorylated by PLK1, 2 and 3, but these enzymes prefer to modify other sites within NFL. PMID: 27503460
fL is a weak independent risk factor in clinically isolated syndromes for conversion to multiple sclerosis. Its role as an axonal damage biomarker may be more relevant as suggested by its association with medium-term brain volume changes. PMID: 27521440
Results from 2 independent prospective cohort studies show that serum NFL is a sensitive and dynamic biomarker for axonal injury in concussive traumatic brain injury. The marker should be useful to detect and monitor CNS injury in concussion PMID: 28404801
Findings support the potential value of serum NfL as a marker of neuroaxonal injury in early multiple sclerosis. Its reduction over time could represent regression to the mean, or a possible treatment effect of IFN-beta-1a. Association with whole brain atrophy and formation of acute white matter lesions has implications to use serum NfL as a biomarker of the overall consequences of brain damage and ongoing disease acti... PMID: 28148632
Higher serum NfL concentrations are associated with more rapid brain atrophy and may therefore reflect disease intensity in dementia (FTD). Because blood sampling is less invasive and has better patient acceptability than lumbar puncture, serum NfL may provide important prognostic information and prove to be a useful outcome measure for clinical trials in FTD. PMID: 27581216
We conclude that the NEFL N98S mutation is associated with a dominant intermediate Charcot-Marie-Tooth disease phenotype characterized by early-onset sensorimotor neuropathy delaying motor milestones, which may evolve into a severe and complex clinical picture including cerebellar ataxia. PMID: 26645395
Plasma Concentration of the Neurofilament Light Protein (NFL) is a Biomarker of CNS Injury in HIV Infection PMID: 26870824
The results showed an important role for miR-25 in regulating NEFL expression in Glioblastoma multiforme. PMID: 26209061
Finally, we demonstrated that NEFL inhibited the NF-kappaB pathway, thereby suppressing the expression of uPA and decreasing NSCLC invasiveness and migration. PMID: 26801564
Cerebrospinal fluid NFL concentration is increased by the early clinical stage of Alzheimer's Disease. PMID: 26524180
The miR-381-NEFL axis is important for temozolomide resistance in glioblastoma multiforme. PMID: 25605243
The results od this study concluded that NEFL E396K mutation may manifest with a novel DI-CMT phenotype, characterized by simultaneous involvement of the peripheral and central nervous system. PMID: 25877835
Suggest monitoring the serum level of antibodies against the NF-L chain as a predictive biomarker of treatment response in multiple sclerosis. PMID: 24515731
a novel heterozygous missense mutation c.1166A>G (p.Y389C) in the gene encoding the light-chain neurofilament protein was identified in 4 patients resulting in hereditary motor and sensory neuropathy with pyramidal signs phenotype. PMID: 25583183
Data suggest that, while lacking a stable structure, NFL-TBS.40-63 peptide (a peptide derived from light neurofilament protein) preferentially binds on a specific single site located near C-terminal end of beta-tubulin. PMID: 26016807
Blood-derived NfL level is an easily accessible biomarker with prognostic value in ALS. PMID: 25934855
Data indicate that the N terminus of Pro-EMAP II binds to its C terminus, arginyl-tRNA synthetase, and the neurofilament light subunit. PMID: 25724651
an NEFL mutation in a family clinically manifesting congenital myopathy PMID: 25264603
NEFL overexpression also enhanced differentiation. PMID: 25312269
This is the first study to suggest blood NFL mRNA as a surrogate marker for early prediction of prediabetic peripheral neuropathy, while NSE mRNA levels may be of no diagnostic value in prediabetic patients. PMID: 24733614
An association was found with high CSF NEFL levels and severity of neurodegenerative diseases and survival. PMID: 25339208
expression of NEFL induces cancer cell apoptosis and inhibits invasion in these cell lines, suggesting that NEFL may play a role in cancer cell apoptosis and invasion. PMID: 23992471
CSF NFL indicates ongoing axonal injury in many neuroasymptomatic HIV patients PMID: 24523921
Our findings reveal an extended phenotype of Charcot Marie Tooth disease 2E caused by an identical missense mutation of the NEFL gene. PMID: 24887401
The expression of two miRNAs (miRNAs miR-b1336 and miR-b2403) whose effect is to stabilize NEFL mRNA, are down regulated in amyotrophic lateral sclerosis. PMID: 24454911
Neurofilament light chain protein NF-L has a dynamic role in the reactive and regenerative changes in axons following injury. PMID: 23802559
Within 7 days after cardiac arrest, serum NF-L is a valuable marker of long-term poor neurological outcome. PMID: 23287695
Increased neurofilament light chain blood levels are associated with neurodegenerative neurological diseases PMID: 24073237
NEFL is downregulated by hypermethylation in breast cancer. PMID: 24026393
CSF neurofilament light chain is elevated in frontotemporal degeneration and correlates with disease severity. PMID: 24242746
The biomarker pattern of neurofilament light in CSF distinguishes between progressive and static neurologic disorders. PMID: 23827424
A group of dysregulated miRNAs directly regulate the NFL mRNA 3'UTR in amyotrophic lateral sclerosis. PMID: 23705811
Anti-NFL antibodies could be surrogate biomarkers of axonal injury in early multiple sclerosis. PMID: 23870535
High CSF NFL levels were found in patients with amyotrophic lateral sclerosis ( PMID: 22680408
Depressed neurofilament expression associates with apolipoprotein E3/E4 genotype in maturing human fetal neurons exposed to HIV-1. PMID: 22302611
NEFL mRNA is ectopically expressed in breast malignancies and could be a potential prognostic factor for early-stage breast cancer patients PMID: 22319610
Identification of gaiting defects combined with previously observed muscle atrophy, reduced axon caliber, and decreased nerve conduction velocity suggests that hNF-L(E397K) mice recapitulate many clinical signs associated with Charcot-Marie-Tooth disease. PMID: 22288874
Cerebrospinal fluid biomarker NF-L in white matter lesions differentiates patients with subcortical vascular disease from those with Alzheimer's. PMID: 21860087
Subjects with APOE epsilon4 have higher CSF NFL levels than non-epsilon4 carriers, only when they do not carry a short poly-T variant of TOMM40, which is associated with later age of onset of AD. PMID: 21983493
Neurofilament light polypeptide (NEFL) was identified as a novel hypermethylated gene associated with resistance to cisplatin-based chemotherapy in cancer of head and neck. PMID: 22246235
Normal cerebrospinal fluid levels of NFL at 12 months post-operatively and beyond suggest the absence of any long-term neuronal damage caused by long-term deep brain stimulation treatment in Parkinson's disease. PMID: 21039366
NFL levels were not significantly increased in multiple sclerosis patients compared with controls and had no relationship to disability or progression PMID: 21197541
While RGNEF and NFL mRNA interact directly in vitro, interestingly they only appear to interact in amyotrophic lateral sclerosis lysates and not in controls PMID: 19488899

Hide All

Involvement in disease

Charcot-Marie-Tooth disease 1F (CMT1F); Charcot-Marie-Tooth disease 2E (CMT2E)

Subcellular Location

Cell projection, axon. Cytoplasm, cytoskeleton.

Protein Families

Intermediate filament family

Database Links

HGNC: 7739

OMIM: 162280

KEGG: hsa:4747

UniGene: Hs.521461

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Code	CSB-EP015688HU
MSDS	MSDS Datasheet
Size	$224
	Order now
Image	(Tris-Glycine gel) Discontinuous SDS-PAGE (reduced) with 5% enrichment gel and 15% separation gel. Based on the SEQUEST from database of E.coli host and target protein, the LC-MS/MS Analysis result of CSB-EP015688HU could indicate that this peptide derived from E.coli-expressed Homo sapiens (Human) NEFL. Based on the SEQUEST from database of E.coli host and target protein, the LC-MS/MS Analysis result of CSB-EP015688HU could indicate that this peptide derived from E.coli-expressed Homo sapiens (Human) NEFL.
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Recombinant Human Neurofilament light polypeptide (NEFL)

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