031402
Kit Type
ELISATests
96Sample Volume
50ulSensitivity
70pg/mlDetection Method
Colorimetric/FluorescentDetection Range
0.156-10ng/mlSample Matrix
serum, plasma, CSFEU Commodity Code
38220000Shipping Temp
Blue IceStorage Temp
4°C/-20°CNeurofilament H, phosphorylated (NF-H, NFH, NEFH) BioAssay™ ELISA Kit
pNF-H
The Neurofilament H, phosphorylated (NF-H, NFH, NEFH) BioAssay™ ELISA Kit measures the levels of phosphorylated NF-H in human, bovine, chicken, equine, mouse and rat serum, plasma and cerebrospinal fluid. Quality control pNF-H is purified from bovine spinal cord. This ELISA assay is intended for research use only.
Principle of the Assay
This ELISA Kit provides a convenient method for measuring level of protein targets within a single, small-volume sample. Neurofilament H, phosphorylated (NF-H, NFH, NEFH) BioAssay™ ELISA Kit is a 2-site sandwich enzyme linked immunoassay providing a plate pre-coated with mouse monoclonal capture antibody against pNF-H. The end-user incubates the samples on the plate and any pNF-H protein in the sample binds to the capture antibody. The end-user then incubates with a solution containing another mouse monoclonal detection antibody conjugated with HRP, which binds to the captured pNF-H. The end-user adds a developer solution that produces a colored product in wells containing HRP, and the intensity of the color corresponds to the amount of pNF-H in the sample. Finally, the end-user stops the reaction by adding H2SO4. The color changes from blue to yellow and can be quantified by measurement on a suitable ELISA plate reader using an absorbance wave length of 450nm.
Kit Components
031402A: Microtiter Strips, 1x96 wells. Precoated with anti-pNF-H 031402B: Detection Antibody (HRP), 4000X , 1x6ul 031402C: Standard (Bovine), 5ug/ml, 1x10ul 031402D: Blocking Protein, 1x500ug 031402E: TBS-Tween Solution, 10X, 1x5ml 031402F: TMB Substrate,1x12ml
Storage and Stability
Store *031402B and *031402C at -20°C. Store all other components at 4°C. Stable for 6 months after receipt. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
Detection Range
0.156-10ng/ml
Precision
Intra-assay CV: <6% Inter-assay CV: <10%
Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.
References
1. Sternberger LA, Sternberger NH. Monoclonal antibodies distinguish phosphorylated and nonphosphorylated forms of neurofilaments in situ. PNAS 80:6126-6130 (1983). 2. Lee VM, Otvos L Jr, Carden MJ, Hollosi M, Dietzschold B, Lazzarini RA. Identification of the major multiphosphorylation site in mammalian neurofilaments. PNAS 85:1998-2002 (1998). 3. Shaw G, Yang C, Ellis R, Anderson K, Parker Mickle J, Scheff S, Pike B, Anderson DK and Howland DR. Hyperphosphorylated neurofilament NF-H is a serum biomarker of axonal injury. Biochem Biophys Res Commun. 336:1268-1277 (2005).4. Petzold, A. and Shaw, G. Comparison of two ELISA methods for measuring levels of the phosphorylated neurofilament heavy chain. J Immunol Methods 319:34-40 (2007).5. Siman, R. et al. Biomarker evidence for mild central nervous system injury after surgically-induced circulation arrest. Brain Res. (2008).6. Lewis, S. et al. Detection of phosphorylated NF-H in the cerebrospinal fluid and blood of aneurysmal subarachnoid hemorrhage patients. J Cereb Blood Flow Metab. Epub Mar 5 (2008). 7. Schlaepfer WW, Lee C, Lee VM, and Zimmerman UJ. An immunoblot study of neurofilament degradation in situ and during calcium-activated proteolysis, J. Neurochem. 44:502–509 (1985). 8. Pant HC. Dephosphorylation of neurofilament proteins enhances their susceptibility to degradation by calpain. Biochem J. 256:665-8 (1988). 9. Stys PK. General mechanism of axonal damage and its prevention. J. Neurol. Sci. 233:3-13 (2005). 10. Buki A and Povlishock JT. All roads lead to disconnection? - Traumatic axonal injury revisited. Acta Neurochir (Wien). 148:181-93 (2005). 11. Matsushige T, Ichiyama T, Anlar B, Tohyama J, Nomura K, Yamashita Y, Furukawa S. CSF neurofilament and soluble TNF receptor 1 levels in subacute sclerosing panencephalitis. J Neuroimmunology 205:155-9] (2008). 12. Boylan, K., Yang, C., Crook, J., Overstreet, K., Heckman, M., Wang, Y., Borchelt, D. and Shaw, G. Immunoreactivity of the phosphorylated axonal neurofilament H subunit (pNF-H) in blood of ALS model rodents and ALS patients: Evaluation of blood pNF-H as a potential ALS biomarker. J. Neurochem 111:1182-1191 (2009).USBio References
No references available