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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.
|Purity||Greater than 90% as determined by SDS-PAGE.|
NEFL; NF68; NFL; Neurofilament light polypeptide; NF-L; 68 kDa neurofilament protein; Neurofilament triplet L protein
|Species||Homo sapiens (Human)|
|Target Protein Sequence||SSFSYEPYYSTSYKRRYVETPRVHISSVRSGYSTARSAYSSYSAPVSSSLSVRRSYSSSSGSLMPSLENLDLSQVAAISNDLKSIRTQEKAQLQDLNDRFASFIERVHELEQQNKVLEAELLVLRQKHSEPSRFRALYEQEIRDLRLAAEDATNEKQALQGEREGLEETLRNLQARYEEEVLSREDAEGRLMEARKGADEAALARAELEKRIDSLMDEISFLKKVHEEEIAELQAQIQYAQISVEMDVTKPDLSAALKDIRAQYEKLAAKNMQNAEEWFKSRFTVLTESAAKNTDAVRAAKDEVSESRRLLKAKTLEIEACRGMNEALEKQLQELEDKQNADISAMQDTINKLENELRTTKSEMARYLKEYQDLLNVKMALDIEIAAYRKLLEGEETRLSFTSVGSITSGYSQSSQVFGRSAYGGLQTSSYLMSTRSFPSYYTSHVQEEQIEVEETIEAAKAEEAKDEPPSEGEAEEEEKDKEEAEEEEAAEEEEAAKEESEEAKEEEEGGEGEEGEETKEAEEEEKKVEGAGEEQAAKKKD
Note: The complete sequence including tag sequence, target protein sequence and linker sequence could be provided upon request.
|Protein Length||Full Length of Mature Protein|
N-terminal 10xHis-SUMO-tagged and C-terminal Myc-tagged
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, pH 8.0.
|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.|
|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.|
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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.
Recombinant Mouse Neurofilament light polypeptide(Nefl)
CSB-YP015688MO >> Yeast
CSB-EP015688MO >> E.coli
CSB-BP015688MO >> Baculovirus
CSB-MP015688MO >> Mammalian cell
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?
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.
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.
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||
|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|