Human Neurofilament protein L (NF-L) ELISA kit

Code CSB-E16094h
Size 96T,5×96T,10×96T
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Product Details

Target Name
neurofilament, light polypeptide
Alternative Names
NEFL; NF68; NFL; Neurofilament light polypeptide; NF-L; 68 kDa neurofilament protein; Neurofilament triplet L protein
Abbreviation
NEFL
Uniprot No.
Species
Homo sapiens (Human)
Sample Types
serum, plasma, tissue homogenates
Detection Range
31.25 pg/mL-2000 pg/mL
Sensitivity
7.8 pg/mL
Assay Time
1-5h
Sample Volume
50-100ul
Detection Wavelength
450 nm
Research Area
Neuroscience
Assay Principle
quantitative
Measurement
Sandwich
Precision
Intra-assay Precision (Precision within an assay): CV%<8%      
Three samples of known concentration were tested twenty times on one plate to assess.  
Inter-assay Precision (Precision between assays): CV%<10%      
Three samples of known concentration were tested in twenty assays to assess.    
             
Linearity
To assess the linearity of the assay, samples were spiked with high concentrations of human NF-L in various matrices and diluted with the Sample Diluent to produce samples with values within the dynamic range of the assay.
  Sample Serum(n=4)  
1:5 Average % 99  
Range % 91-105  
1:10 Average % 85  
Range % 80-92  
1:20 Average % 88  
Range % 82-94  
1:40 Average % 93  
Range % 86-98  
Recovery
The recovery of human NF-L spiked to levels throughout the range of the assay in various matrices was evaluated. Samples were diluted prior to assay as directed in the Sample Preparation section.
Sample Type Average % Recovery Range  
Serum (n=5) 95 89-98  
EDTA plasma (n=4) 98 92-104  
             
             
Typical Data
These standard curves are provided for demonstration only. A standard curve should be generated for each set of samples assayed.
pg/ml OD1 OD2 Average Corrected  
2000 2.770 2.665 2.718 2.628  
1000 2.114 2.006 2.060 1.970  
500 1.256 1.243 1.250 1.160  
250 0.657 0.651 0.654 0.564  
125 0.343 0.339 0.341 0.251  
62.5 0.252 0.242 0.247 0.157  
31.25 0.209 0.198 0.204 0.114  
0 0.092 0.088 0.090    
Materials provided
  • A micro ELISA plate ---The 96-well plate has been pre-coated with an anti-human NEFL antibody. This dismountable microplate can be divided into 12 x 8 strip plates.
  • Two vials lyophilized standard ---Dilute a bottle of the standard at dilution series, read the OD values, and then draw a standard curve.
  • One vial Biotin-labeled NEFL antibody (100 x concentrate) (120 μl/bottle) ---Act as the detection antibody.
  • One vial HRP-avidin (100 x concentrate) (120 μl/bottle) ---Bind to the detection antibody and react with the TMB substrate to make the solution chromogenic.
  • One vial Biotin-antibody Diluent (15 ml/bottle) ---Dilute the Biotin-antibody.
  • One vial HRP-avidin Diluent (15 ml/bottle) ---Dilute the HRP-avidin solution.
  • One vial Sample Diluent (50 ml/bottle)---Dilute the sample to an appropriate concentration.
  • One vial Wash Buffer (25 x concentrate) (20 ml/bottle) ---Wash away unbound or free substances.
  • One vial TMB Substrate (10 ml/bottle) ---Act as the chromogenic agent. TMB interacts with HRP, eliciting the solution turns blue.
  • One vial Stop Solution (10 ml/bottle) ---Stop the color reaction. The solution color immediately turns from blue to yellow.
  • Four Adhesive Strips (For 96 wells) --- Cover the microplate when incubation.
  • An instruction manual
Materials not provided
  • A microplate reader capable of measuring absorbance at 450 nm, with the correction wavelength set at 540 nm or 570 nm.
  • An incubator can provide stable incubation conditions up to 37°C±5°C.
  • Centrifuge
  • Vortex
  • Squirt bottle, manifold dispenser, or automated microplate washer
  • Absorbent paper for blotting the microtiter plate
  • 50-300ul multi-channel micropipette
  • Pipette tips
  • Single-channel micropipette with different ranges
  • 100ml and 500ml graduated cylinders
  • Deionized or distilled water
  • Timer
  • Test tubes for dilution
Troubleshooting
and FAQs
Storage
Store at 2-8°C. Please refer to protocol.
Lead Time
3-5 working days after you place the order, and it takes another 3-5 days for delivery via DHL or FedEx
Description

This Human NEFL ELISA Kit was designed for the quantitative measurement of Human NEFL protein in serum, plasma, tissue homogenates. It is a Sandwich ELISA kit, its detection range is 31.25 pg/mL-2000 pg/mL and the sensitivity is 7.8 pg/mL.

Customer Reviews and Q&A

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 Q&A
Q:

The Cusabio ELISA kit CSB-E16094h, does not name the source of the standard. Can you tell us which standard is used here?
Could you send us linearity and parallelism data for this ELISA?

A:
Thanks for your inquiry.
The standard of this kit is 293 cell-derived recombinant protein.
As for the linearity and parallelism data,I will send it to you via e-mail.Pls let me know if you have any further questions. Thank you.

Target Background

Function
(From Uniprot)
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
  1. NF-L may be involved in severity of neuronal injury following traumatic brain injury PMID: 27819296
  2. CSF neurofilament light chain protein is an accurate marker for distinguishing Alzheimer's disease patients from healthy controls. PMID: 28527630
  3. Cerebrospinal fluid neurofilament protein light is a useful tool for determining disease intensity in frontotemporal dementia and motor neuron disease patients. PMID: 28631955
  4. 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
  5. In this Chinese Han population a novel Charcot-Marie-Tooth disease-associated gene mutations of NEFL (c.280C>T) was discovered. PMID: 27862672
  6. 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
  7. 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
  8. 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
  9. 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
  10. 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
  11. 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
  12. 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
  13. 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
  14. Plasma Concentration of the Neurofilament Light Protein (NFL) is a Biomarker of CNS Injury in HIV Infection PMID: 26870824
  15. The results showed an important role for miR-25 in regulating NEFL expression in Glioblastoma multiforme. PMID: 26209061
  16. Finally, we demonstrated that NEFL inhibited the NF-kappaB pathway, thereby suppressing the expression of uPA and decreasing NSCLC invasiveness and migration. PMID: 26801564
  17. Cerebrospinal fluid NFL concentration is increased by the early clinical stage of Alzheimer's Disease. PMID: 26524180
  18. The miR-381-NEFL axis is important for temozolomide resistance in glioblastoma multiforme. PMID: 25605243
  19. 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
  20. Suggest monitoring the serum level of antibodies against the NF-L chain as a predictive biomarker of treatment response in multiple sclerosis. PMID: 24515731
  21. 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
  22. 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
  23. Blood-derived NfL level is an easily accessible biomarker with prognostic value in ALS. PMID: 25934855
  24. 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
  25. an NEFL mutation in a family clinically manifesting congenital myopathy PMID: 25264603
  26. NEFL overexpression also enhanced differentiation. PMID: 25312269
  27. 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
  28. An association was found with high CSF NEFL levels and severity of neurodegenerative diseases and survival. PMID: 25339208
  29. 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
  30. CSF NFL indicates ongoing axonal injury in many neuroasymptomatic HIV patients PMID: 24523921
  31. Our findings reveal an extended phenotype of Charcot Marie Tooth disease 2E caused by an identical missense mutation of the NEFL gene. PMID: 24887401
  32. 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
  33. Neurofilament light chain protein NF-L has a dynamic role in the reactive and regenerative changes in axons following injury. PMID: 23802559
  34. Within 7 days after cardiac arrest, serum NF-L is a valuable marker of long-term poor neurological outcome. PMID: 23287695
  35. Increased neurofilament light chain blood levels are associated with neurodegenerative neurological diseases PMID: 24073237
  36. NEFL is downregulated by hypermethylation in breast cancer. PMID: 24026393
  37. CSF neurofilament light chain is elevated in frontotemporal degeneration and correlates with disease severity. PMID: 24242746
  38. The biomarker pattern of neurofilament light in CSF distinguishes between progressive and static neurologic disorders. PMID: 23827424
  39. A group of dysregulated miRNAs directly regulate the NFL mRNA 3'UTR in amyotrophic lateral sclerosis. PMID: 23705811
  40. Anti-NFL antibodies could be surrogate biomarkers of axonal injury in early multiple sclerosis. PMID: 23870535
  41. High CSF NFL levels were found in patients with amyotrophic lateral sclerosis ( PMID: 22680408
  42. Depressed neurofilament expression associates with apolipoprotein E3/E4 genotype in maturing human fetal neurons exposed to HIV-1. PMID: 22302611
  43. NEFL mRNA is ectopically expressed in breast malignancies and could be a potential prognostic factor for early-stage breast cancer patients PMID: 22319610
  44. 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
  45. Cerebrospinal fluid biomarker NF-L in white matter lesions differentiates patients with subcortical vascular disease from those with Alzheimer's. PMID: 21860087
  46. 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
  47. 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
  48. 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
  49. NFL levels were not significantly increased in multiple sclerosis patients compared with controls and had no relationship to disability or progression PMID: 21197541
  50. 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

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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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