Human Neurofilament protein H (NF-H) ELISA kit

Code CSB-E16097h
Size 96T,5×96T,10×96T
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Product Details

Target Name
neurofilament, heavy polypeptide
Alternative Names
200 kDa neurofilament protein ELISA Kit; CMT2CC ELISA Kit; Nefh ELISA Kit; Neurofilament heavy polypeptide 200kDa ELISA Kit; Neurofilament heavy polypeptide ELISA Kit; Neurofilament triplet H protein ELISA Kit; NF H ELISA Kit; NF-H ELISA Kit; NFH ELISA Kit; NFH_HUMAN ELISA Kit
Abbreviation
NEFH
Uniprot No.
Species
Homo sapiens (Human)
Sample Types
serum, plasma, cerebrospinal fluid (CSF)
Detection Range
0.625 ng/mL-40 ng/mL
Sensitivity
0.156 ng/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-H in various matrices and diluted with the Sample Diluent to produce samples with values within the dynamic range of the assay.
SampleSerum(n=4)
1:1Average %92
Range %86-98
1:2Average %97
Range %93-101
1:4Average %93
Range %84-97
1:8Average %103
Range %95-107
Recovery
The recovery of Human NF-H 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 TypeAverage % RecoveryRange
Serum (n=5) 8580-90
EDTA plasma (n=4)9488-98
Typical Data
These standard curves are provided for demonstration only. A standard curve should be generated for each set of samples assayed.
ng/mlOD1OD2AverageCorrected
402.513 2.614 2.564 2.396
202.017 2.115 2.066 1.898
101.442 1.343 1.393 1.225
50.887 0.864 0.876 0.708
2.50.531 0.545 0.538 0.370
1.250.374 0.362 0.368 0.200
0.6250.276 0.284 0.280 0.112
00.167 0.168 0.168
Materials provided
  • A micro ELISA plate --- The 96-well plate has been pre-coated with an anti-human NF-H 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 NF-H 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-antibodyDiluent (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 NEFH ELISA Kit was designed for the quantitative measurement of Human NEFH protein in serum, plasma, cerebrospinal fluid (CSF). It is a Sandwich ELISA kit, its detection range is 0.625 ng/mL-40 ng/mL and the sensitivity is 0.156 ng/mL.

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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. NEFH has an important function in mature axons that is not subserved by the two smaller NF proteins. May additionally cooperate with the neuronal intermediate filament proteins PRPH and INA to form neuronal filamentous networks.
Gene References into Functions
  1. In newborns undergoing cardiac surgery, phosphorylated axonal neurofilament heavy chain was decreased at 0 hours in both the cardiopulmonary bypass and deep hypothermic circulatory arrest groups compared to baseline. PMID: 29945509
  2. Phosphoneurofilament heavy chain level was higher in ALS patients than in controls PMID: 27538346
  3. Data suggest that high CSF NfH levels are an early predictor of later brain and spinal cord atrophy in multiple sclerosis patients. PMID: 27207456
  4. Findings in the dorsolateral prefrontal cortex in schizophrenia provide evidence of altered proteins involved in synaptic function (FABP4), cytoarchitecture organization (NEFH), and circadian molecular clock signaling (CSNK1E), which may be contributing to the cognitive and/or negative symptoms in this disorder. FABP4, CSNK1E and NEFH could become potentially useful biomarkers for schizophrenia. PMID: 27236410
  5. Unique deletions of two nucleotides causing frameshifts near the end of the NEFH coding sequence were identified in two Charcot-Marie-Tooth disease families. Using electroporation of chick embryo spinal cord, confirmed that NEFH mutants form aggregates in vivo and trigger apoptosis of spinal cord neurons. PMID: 28709447
  6. This study confirmed the general applicability of the monocentric obtained cut-off values for neurofilamens in ALS, especially for pNfH PMID: 27415180
  7. data support the use of Cebrospinal fluid phosphorylated NFH as a prognostic biomarker for amyotrophic lateral sclerosis. PMID: 28628244
  8. Study found a significant correlation between values of 8-hydroxy-2'-deoxyguanosine and phosphorylated NF-H only in clinically isolated syndrome group. While the plasma values of 8-hydroxy-2'-deoxyguanosine reflect the degree of acute demyelination in clinically isolated syndrome, phosphorylated NF-H values reflect that in relapsing-remitting multiple sclerosis. PMID: 27295058
  9. Results provide evidence that in particular pNfH can be used as a good diagnostic biomarker of ALS at the diagnostic stage. Moreover, results indicate that NfL may be useful in monitoring disease progression in a subset of patients. PMID: 28500227
  10. Level of neurofilament heavy chain and light chains were significantly elevated in the cerebrospinal fluid of Amyotrophic Lateral Sclerosis (ALS) patients compared to healthy controls/controls without parenchymal central nervous system involvement and ALS mimic disease patients. PMID: 27732645
  11. Study found a significant increase of pNF-H levels in both plasma and CSF in amyotrophic lateral sclerosis patients PMID: 27423602
  12. Studied diagnostic Value of Serum Levels of GFAP, pNF-H, and NSE Compared With Clinical Findings in Severity Assessment of Human Traumatic Spinal Cord Injury. PMID: 25341992
  13. This study demonstrated that Higher NF-L concentrations (beta = -0.26) were associated with functional decline in patients with vascular burden. PMID: 25633679
  14. Phospho-NFH levels were significantly higher in amyotrophic lateral sclerosis patients in comparison with controls, in particular in fast progressors. PMID: 25261856
  15. Data identified NEFH methylation as a candidate epigenetic marker for prognosis of RCC patients as well as prediction of anti-vascular endothelial growth factor-based therapy response. PMID: 24464810
  16. pNFL-H may be useful in determining which individuals require CT imaging to assess the severity of their injury PMID: 25192482
  17. Subconcussive repetitive trauma in amateur boxing causes a mild traumatic brain injury that may be diagnosed by CSF analysis of expressed pNFH, even without unconsciousness or concussion symptoms. PMID: 24260563
  18. Absence of axonal neurofilaments in NFH-LacZ transgenic mice compromises axonal regeneration. PMID: 23079625
  19. In conclusion, pNfH represents a promising candidate for inclusion in a panel of diagnostic and prognostic biomarkers. PMID: 23134506
  20. CSF and intrathecal levels and CSF/serum ratios of anti-NFH antibodies were increased in the CIS patients early developing multiple sclerosis. PMID: 23632043
  21. These data support further study of pNF-H in CSF, serum and plasma as a potential amyotrophic lateral sclerosis biomarker PMID: 23117489
  22. NF-H levels increase significantly faster in children who have a worse Glasgow Outcome Scale following traumatic brain injury, or died. PMID: 21976236
  23. significant in vivo information on the pathophysiology of stroke may be obtained by the determination of NfH(SMI35). PMID: 21792676
  24. Serum neurofilament protein H levels were significantly elevated in stroke patients compared to healthy controls. PMID: 21349546
  25. Data suggest that a deficiency of cellular neuroprotective mechanisms (decrease of sAPP) is linked to progressive neuro-axonal damage (increase of NfH(SMI35)) and to progression of disease. PMID: 21858182
  26. Data show that RNA interference-mediated knockdown of NEFH accelerated ESCC cell growth in culture and increased tumorigenicity in vivo. PMID: 20140245
  27. NEFH is phosphorylated at serine 493 by GSK3b PMID: 12130654
  28. NEFH gene is involved in the pathogenesis of sporadic motor neuron disease PMID: 14722583
  29. mutations in neurofilaments are possible risk factors that may contribute to pathogenesis in amyotrophic lateral sclerosis in conjunction with one or more additional genetic or environmental factors, but are not significant primary causes PMID: 16084104
  30. A subgroup of FTD patients had remarkably high CSF levels of NfH. The degree of NfH phosphorylation was increased in FTD compared to both other groups. PMID: 17290105
  31. Isomerization of lys-ser-pro repeat residues that are abundant in NF-H tail domains by Pin1 can regulate NF-H phosphorylation. PMID: 17626162
  32. analysis of major neurofilament subunit NF-H levels in blood and cerebrospinal fluid differentiates between patients with poor and favorable outcomes. PMID: 18319731
  33. Pin1 as a possible modulator of stress-induced NF-H phosphorylation as seen in neurodegenerative disorders like AD and amyotrophic lateral sclerosis PMID: 18635547
  34. Elevated pNF-H released into the serum of some affected Leber hereditary optic neuropathy patients may suggest that axonal degeneration occurs at some point after loss of visual function. PMID: 19104679
  35. cerebrospinal fluid phosphorylated forms of neurofilament heavy subunit are not molecular markers of axonal damage for HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP) probably due to the slow progression of this disease. PMID: 19678766

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Involvement in disease
Amyotrophic lateral sclerosis (ALS); Charcot-Marie-Tooth disease 2CC (CMT2CC)
Subcellular Location
Cytoplasm, cytoskeleton. Cell projection, axon.
Protein Families
Intermediate filament family
Database Links

HGNC: 7737

OMIM: 105400

KEGG: hsa:4744

STRING: 9606.ENSP00000311997

UniGene: Hs.198760

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