Human Neurofilament H ELISA Kit (200 kDa Neurofilament protein) (ab272199)
Key features and details
- One-wash 90 minute protocol
- Sensitivity: 141 pg/ml
- Range: 0.94 ng/ml - 60 ng/ml
- Sample type: Cell Lysate, Tissue Extracts
- Detection method: Colorimetric
- Assay type: Sandwich (quantitative)
- Reacts with: Human
Overview
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Product name
Human Neurofilament H ELISA Kit (200 kDa Neurofilament protein)
See all Neurofilament heavy polypeptide kits -
Detection method
Colorimetric -
Precision
Intra-assay Sample n Mean SD CV% Extract 8 6% Inter-assay Sample n Mean SD CV% Extract 3 2.1% -
Sample type
Tissue Extracts, Cell Lysate -
Assay type
Sandwich (quantitative) -
Sensitivity
141 pg/ml -
Range
0.94 ng/ml - 60 ng/ml -
Recovery
Sample specific recovery Sample type Average % Range Tissue Extracts 120 115% - 127% Cell Lysate 86 82% - 90% -
Assay time
1h 30m -
Assay duration
One step assay -
Species reactivity
Reacts with: Human -
Product overview
Human Neurofilament H ELISA kit (ab272199) is a single-wash 90 min sandwich ELISA designed for the quantitative measurement of Human Neurofilament H protein in human cell and tissue extract samples. It uses our proprietary SimpleStep ELISA® technology. Quantitate Human Neurofilament H with 1.41 pg/mL sensitivity.
SimpleStep ELISA® technology employs capture antibodies conjugated to an affinity tag that is recognized by the monoclonal antibody used to coat our SimpleStep ELISA® plates. This approach to sandwich ELISA allows the formation of the antibody-analyte sandwich complex in a single step, significantly reducing assay time. See the SimpleStep ELISA® protocol summary in the image section for further details. Our SimpleStep ELISA® technology provides several benefits:
-Single-wash protocol reduces assay time to 90 minutes or less
-High sensitivity, specificity and reproducibility from superior antibodies
-Fully validated in biological samples
-96-wells plate breakable into 12 x 8 wells stripsA 384-well SimpleStep ELISA® microplate (ab203359) is available to use as an alternative to the 96-well microplate provided with SimpleStep ELISA® kits.
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Notes
Neurofilament H, also known as 200 kDa Neurofilament Protein, together with Neurofilament L and M are three intermediate filament proteins of neurofilaments. Neurofilament H is involved in the maintenance of neuronal caliber and may play a role in intracellular transport. Neurofilament H is used as a neuronal damage biomarker and mutations to the gene have been associated with susceptibility to amyotrophic lateral sclerosis. The human Neurofilament H amino acids sequence is 75% identical to mouse and rat Neurofilament H sequence.
Abcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses. -
Platform
Pre-coated microplate (12 x 8 well strips)
Properties
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Storage instructions
Store at +4°C. Please refer to protocols. -
Components 1 x 96 tests 10X Human Neurofilament H (200 kDa Neurofilament Protein) Capture Antibody 1 x 600µl 10X Human Neurofilament H (200 kDa Neurofilament Protein) Detector Antibody 1 x 600µl 10X Wash Buffer PT (ab206977) 1 x 20ml 50X Cell Extraction Enhancer Solution (ab193971) 1 x 1ml 5X Cell Extraction Buffer PTR (ab193970) 1 x 10ml Antibody Diluent 5BI 1 x 6ml Human Neurofilament H (200 kDa Neurofilament Protein) Lyophilized Recombinant Protein 2 vials Plate Seals 1 unit Sample Diluent NS (ab193972) 1 x 12ml SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit Stop Solution 1 x 12ml TMB Development Solution 1 x 12ml -
Research areas
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Function
Neurofilaments usually contain three intermediate filament proteins: L, M, and H which are involved in the maintenance of neuronal caliber. NF-H has an important function in mature axons that is not subserved by the two smaller NF proteins. -
Involvement in disease
Defects in NEFH are a cause of susceptibility to amyotrophic lateral sclerosis (ALS) [MIM:105400]. ALS is a neurodegenerative disorder affecting upper and lower motor neurons, and resulting in fatal paralysis. Sensory abnormalities are absent. Death usually occurs within 2 to 5 years. The etiology is likely to be multifactorial, involving both genetic and environmental factors. -
Sequence similarities
Belongs to the intermediate filament family. -
Post-translational
modificationsThere are a number of repeats of the tripeptide K-S-P, NFH is phosphorylated on a number of the serines in this motif. It is thought that phosphorylation of NFH results in the formation of interfilament cross bridges that are important in the maintenance of axonal caliber.
Phosphorylation seems to play a major role in the functioning of the larger neurofilament polypeptides (NF-M and NF-H), the levels of phosphorylation being altered developmentally and coincident with a change in the neurofilament function.
Phosphorylated in the Head and Rod regions by the PKC kinase PKN1, leading to inhibit polymerization. - Information by UniProt
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Alternative names
- 200 kDa neurofilament protein
- CMT2CC
- Nefh
see all -
Database links
- Entrez Gene: 4744 Human
- Omim: 162230 Human
- SwissProt: P12036 Human
- Unigene: 198760 Human
Images
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SimpleStep ELISA Protocol Diagram
SimpleStep ELISA technology allows the formation of the antibody-antigen complex in one single step, reducing assay time to 90 minutes. Add samples or standards and antibody mix to wells all at once, incubate, wash, and add your final substrate. See protocol for a detailed step-by-step guide.
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Example of human Neurofilament H standard curve in 1X Cell Extraction Buffer PTR + Enhancer.The Neurofilament H standard curve was prepared as described in Section 10. Raw data values are shown in the table. Background-subtracted data values (mean +/- SD) are graphed.
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Interpolated concentrations of native Neurofilament H in human SH-SY5Y cell extract based on a 250 µg/mL extract load and human brain tissue extract based on a 10 µg/mL extract load.The concentrations of Neurofilament H were measured in duplicate and interpolated from the Neurofilament H standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Neurofilament H concentration was determined to be 22.94 ng/mL in SH-SY5Y cell extract and 57.07 ng/mL in human brain tissue extract.
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Species reactivity was determined by measuring the protein in various samples, interpolating the protein concentrations from the human standard curve, and expressing the interpolated concentrations.The mean Neurofilament H concentration was determined to be 5.301 µg/mg in human brain tissue extract, 3.386 µg/mg in mouse brain tissue extract, and 0.216 µg/mg in rat brain tissue extract.
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Sandwich ELISA - Human Neurofilament H ELISA Kit (200 kDa Neurofilament protein) (ab272199)To learn more about the advantages of recombinant antibodies see here.
