Purified anti-Neurofilament H (NF-H), Phosphorylated Antibody (Previously Covance catalog# SMI 31P)

Pricing & Availability
Clone
SMI 31 (See other available formats)
Regulatory Status
RUO
Other Names
Neurofilament heavy polypeptide, NF-H, 200 kD neurofilament protein, neurofilament triplet H protein
Previously
Covance Catalog# SMI 31P
Isotype
Mouse IgG1, κ
A_SMI-31_PURE_NF-H_Antibody_IHCP_Mouse_011718
IHC staining of purified anti-Neurofilament H (NF-H), Phosphorylated antibody (clone SMI 31) on formalin-fixed paraffin-embedded mouse brain tissue. Following antigen retrieval using Retrieval-ALL Antigen Unmasking System 3 (Cat. No. 927601), the tissue was incubated with 1 µg/ml of the primary antibody overnight at 4°C. BioLegend’s Ultra Streptavidin (USA) HRP Detection Kit (Multi-Species, DAB, Cat. No. 929901) was used for detection followed by hematoxylin counterstaining, according to the protocol provided. The image was captured with a 40X objective.
  • A_SMI-31_PURE_NF-H_Antibody_IHCP_Mouse_011718
    IHC staining of purified anti-Neurofilament H (NF-H), Phosphorylated antibody (clone SMI 31) on formalin-fixed paraffin-embedded mouse brain tissue. Following antigen retrieval using Retrieval-ALL Antigen Unmasking System 3 (Cat. No. 927601), the tissue was incubated with 1 µg/ml of the primary antibody overnight at 4°C. BioLegend’s Ultra Streptavidin (USA) HRP Detection Kit (Multi-Species, DAB, Cat. No. 929901) was used for detection followed by hematoxylin counterstaining, according to the protocol provided. The image was captured with a 40X objective.
  • B_SMI-31_PURE_NF-H_Antibody_IHCP_Rat_011718
    IHC staining of purified anti-Neurofilament H (NF-H), Phosphorylated antibody (clone SMI 31) on formalin-fixed paraffin-embedded rat brain tissue. After antigen retrieval using Retrieval-ALL Antigen Unmasking System 3 (Cat. No. 927601), the tissue was incubated with 5 µg/mL of the primary antibody overnight at 4°C followed by incubation with 2.5 µg/ml of Alexa Fluor® 594 goat anti-mouse IgG for one hour at room temperature. The image was captured with a 40X objective.
  • C_SMI-31_PURE_NF-H_Antibody_WB_011718
    Western blot of purified anti-Neurofilament H (NF-H), Phosphorylated antibody (clone SMI 31). Lane 1: Molecular weight marker; Lane 2: 20 µg of human brain lysate; Lane 3: 20 µg of mouse brain lysate; Lane 4: 20 µg of rat brain lysate. The blots were incubated with 1 µg/mL of SMI 31 or mouse IgG1 overnight at 4°C, followed by incubation with HRP-labeled goat anti-mouse IgG (Cat. No. 405306). Direct-Blot™ HRP anti-Tubulin Beta 3 (TUBB3) antibody (clone AA10, Cat. No. 657409) was used as a loading control. Enhanced chemiluminescence was used as the detection system.
  • D_SMI-31_PURE_NF-H_Antibody_IHCP_Rat2_011718
    IHC staining of purified anti-Neurofilament H (NF-H), Phosphorylated antibody (clone SMI 31) on formalin-fixed paraffin-embedded rat brain tissue. Following antigen retrieval using Retrieval-ALL Antigen Unmasking System 3 (Cat. No. 927601), the tissue was incubated with 1 µg/ml of the primary antibody overnight at 4°C. BioLegend´s Ultra Streptavidin (USA) HRP Detection Kit (Multi-Species, DAB, Cat. No. 929901) was used for detection followed by hematoxylin counterstaining, according to the protocol provided. The image was captured with a 40X objective.
  • E_SMI31_NeurofilamentH_Phosphorylated_ICC_012918
    ICC staining of purified anti-Neurofilament H (NF-H), Phosphorylated antibody (clone SMI 31) on SH-SY5Y neuroblastoma cells. The cells were fixed with 4% PFA, permeabilized with a buffer containing 0.1% Triton X-100 and 0.25% BSA, and blocked with 2% normal goat serum and 0.02% BSA. The cells were then incubated with 1 µg/ml of the primary antibody overnight at 4°C, followed by incubation with 2.5 µg/ml of Alexa Fluor® 594 goat anti-mouse IgG for one hour at room temperature. The image was captured with a 40X objective 0.02% BSA. The cells were then incubated with 1 µg/ml of the primary antibody overnight at 4°C, followed by incubation with 2.5 µg/ml of Alexa Fluor® 594 goat anti-mouse IgG for one hour at room temperature. The image was captured with a 40X objective.
  • F_SMI-31_PURE_NF-H_Antibody_IHCP_Mouse2_011718
    Paraformaldehyde-perfused GFP mouse cerebellum was permeabilized with PBS-Triton X (0.4%) for 30 minutes, and heat-induced antigen retrieval was performed with pH 6.0 sodium citrate. Slides were blocked with 10% donor horse serum for one hour, then stained with 0.5 - 2 µg/mL of purified anti-Neurofilament H (NF-H), Phosphorylated antibody (clone SMI 31, red) along with NF200 monoclonal antibody (blue). After washing, sections were stained with 1:200 secondary antibody. Credit: Dr. Michael R Williams, Luikart Lab, Dartmouth College.
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801602 25 µL $112.00
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801601 100 µL $276.00
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Description

Neurofilaments (NF) are approximately 10 nanometer intermediate filaments found in neurons. They are a major component of the neuronal cytoskeleton, and function primarily to provide structural support for the axon and to regulate the axon diameter. There are three major NF subunits, and the names given to these subunits are based upon the apparent molecular mass of the mammalian subunits on SDS-PAGE. The light or lowest NF (NF-L) runs at 68-70 kD. The medium or middle NF (NF-M) runs at about 145-160 kD, and the heavy or highest NF (NF-H) runs at 200-220 kD. However, the actual molecular weight of these proteins is considerably lower due to the highly charged C-terminal regions of the molecules. The level of NF gene expression correlates with the axonal diameter, which controls how fast electrical signals travel down the axon. Mutant mice with NF abnormalities have phenotypes resembling amyotrophic lateral sclerosis. NF immunostaining is common in diagnostic neuropathology. It is useful for differentiating neurons (positive for NF) from the glia (negative for NF).

Technical data sheet

Product Details

Verified Reactivity
Human, Mouse, Rat
Antibody Type
Monoclonal
Host Species
Mouse
Formulation
Phosphate-buffered solution + Thimerosal.
Preparation
The antibody was purified by affinity chromatography.
Concentration
1 mg/ml
Storage & Handling
The antibody solution should be stored undiluted between 2°C and 8°C. Please note the storage condition for this antibody has been changed from -20°C to between 2°C and 8°C. You can also check your vial or your CoA to find the most accurate storage condition for this antibody.
Application

IHC-P - Quality tested
WB, ICC - Verified
Recommended Usage

Each lot of this antibody is quality control tested by formalin-fixed paraffin-embedded immunohistochemical staining. For immunohistochemistry, a concentration range of 1.0 - 5.0 µg/ml is suggested. For Western blotting, the suggested use of this reagent is 1.0 - 5.0 µg/ml. For immunocytochemistry, the suggested use of this reagent is 1.0 - 5.0 µg/ml. It is recommended that the reagent be titrated for optimal performance for each application.

Application Notes

Additional reported applications (for the relevant formats) include: Western blotting1, immunohistochemistry2,4, and immunocytochemistry4.

SMI 31 reacts with a phosphorylated epitope in extensively phosphorylated neurofilament H and, to a lesser extent, with neurofilament M in most mammalian species, which chicken and frog (Xenopus). Immunocytochemically, SMI 31 reacts broadly with thick and thin axons and some dendrites such as basket cell dendrites, but not Purkinje cell dendrites. Nerve cell bodies are generally unreactive. Other cells and tissues are unreactive except for peripheral axons. Phosphatase treatment of tissue sections or Western blots abolishes reaction with SMI 31. Staining is unaffected by trypsin. In pathological conditions, reaction with SMI 31 may be found also in neuronal cell bodies. Aberrant phosphorylation of neurofilament H in cell bodies can be demonstrated in neuronal cell cultures with SMI 31 by agents that induce stress-activated protein kinase. In its reaction with paired helical filaments in hereditary inclusion body myopathy, SMI 31 colocalizes with nitric oxide synthase, suggesting that oxidative stress may play a role in the pathogenic cascade of such degenerative diseases. SMI 31 co-immunoprecipitates neurofilament-associated kinase (NAK 115) via reaction of the antibody with the tail domain of neurofilament H.

Application References

(PubMed link indicates BioLegend citation)
  1. Barry D, et al. 2012. J. Neurosci. 32:6209 (WB) PubMed
  2. Choi Y, et al. 2008. Genes Dev. 22:2485. (IHC) PubMed
  3. Sepulveda B, et al. 2013. PLoS ONE. 8(e61986. (ICC) PubMed
  4. McLean NA, et. al. 2014. PLoS One 9:e110174. (IHC) PubMed
Product Citations
  1. Fleuriet J, et al. 2020. Sci Rep. 10:11927. PubMed
  2. Seiler S, et al. 2023. Sci Rep. 13:2883. PubMed
  3. D'mello V, et al. 2023. Neurotrauma Rep. 4:236. PubMed
  4. So HK, et al. 2023. Research (Wash D C). 6:0158. PubMed
  5. Frondelli MJ, et al. 2022. J Neurosci Res. 100:578. PubMed
  6. Powers RM, et al. 2022. Mol Biol Cell. 33:ar64. PubMed
  7. Cunningham ME 2022. Experimental Neurology. 355:114127. PubMed
  8. Won D, et al. 2022. Sci Adv. 8:eabo3209. PubMed
  9. Hu T, et al. 2022. Cell Biosci. 12:196. PubMed
  10. Pellegatta M, et al. 2022. J Neurosci. 42:2433. PubMed
  11. Zhu PP, et al. 2022. Hum Mol Genet. 31:2779. PubMed
  12. Takahashi M, et al. 2015. J Neurol Neurosurg Psychiatry. 86: 939-944. PubMed
  13. Johari M, et al. 2021. Acta Neuropathologica. . PubMed
  14. Lavoie–Cardinal F, et al. 2020. Sci Rep. 10:11960. PubMed
  15. von Saucken VE, et al. 2020. Neurobiol Dis. 145:105072. PubMed
  16. Miguel–Hidalgo JJ, et al. 2019. Neuroscience. 411:255. PubMed
  17. McGonigal R, et al. 2021. Journal of Neurochemistry. . PubMed
  18. Alada-Morais S, et al. 2021. Cerebral Cortex. :. PubMed
  19. Wang S et al. 2019. Front Cell Neurosci. 0.766666667 . PubMed
  20. Mahajan KR, et al. 2020. Ann Neurol. 88:81. PubMed
  21. Andrew RJ, et al. 2017. Proc Natl Acad Sci U S A. 114:E9665. PubMed
  22. Özen I, et al. 2022. Acta Neuropathol Commun. 10:129. PubMed
  23. Cui X, et al. 2016. Stroke. 47: 214 - 220. PubMed
  24. Berghoff S, et al. 2017. Nat Commun. 8:14241. PubMed
  25. Barry D, et al. 2012. J Neurosci. 32:6209-6219. PubMed
  26. Cui C, et al. 2016. Stem Cells Trans Med. 5(12):1656-1667. PubMed
  27. Wang W, et al. 2022. Dose Response. 20:15593258221112959. PubMed
  28. McGonigal R, et al. 2022. J Clin Invest. :. PubMed
  29. Manangeeswaran M, et al. 2022. Front Immunol. 13:919815. PubMed
  30. Choi Y, et al. 2008. Genes Dev. 22:2485-2495. PubMed
  31. Maggiore JC, et al. 2020. J Tissue Eng Regen Med. 14:1892. PubMed
  32. Xu X, et al. 2022. Sci Rep. 12:14690. PubMed
  33. Stumpf SK, et al. 2019. Acta Neuropathol. 138:147. PubMed
  34. Scaramuzzino C, et al. 2022. Sci Adv. 8:eabj8812. PubMed
  35. McGonigal R et al. 2018. The Journal of Neuroscience. 39(1):63-77 . PubMed
  36. Chaves RS, et al. 2021. J Neurosci. 41:10034. PubMed
  37. Maire C, et al. 2014. Stem Cells. 32:313-326. PubMed
  38. Redondo J, et al. 2015. Brain Pathol. 25:692-700. PubMed
  39. Generous AR, et al. 2019. J Cell Sci. 132:jcs235507. PubMed
  40. Cuadrado E et al. 2019. Cell reports. 26(7):1718-1726 . PubMed
  41. Halabi EA, et al. 2019. Nat Commun. 10:1232. PubMed
  42. Yu G, et al. 2018. Neuroreport. 28:29. PubMed
  43. McLean N, et al. 2014. PLoS One. 9:e110174. PubMed
  44. Riku Y, et al. 2016. J Neuropathol Exp Neurol. 75: 801 - 811. PubMed
  45. Kubo A, et al. 2019. J Comp Neurol. 527:985. PubMed
  46. Bishop R, et al. 2022. Sci Rep. 12:143. PubMed
  47. Kiryu-Seo S, et al. 2010. J Neurosci. 30:6658-6666. PubMed
  48. Schultz V, et al. 2021. Glia. 69:2023. PubMed
  49. Doroshenko ER, et al. 2021. Front Immunol. 570425:12. PubMed
  50. Cooper ML, et al. 2020. Proc Natl Acad Sci U S A. 117:18810. PubMed
  51. Li T, et al. 2016. Nat Commun. 7:12082. PubMed
RRID
AB_2564641 (BioLegend Cat. No. 801602)
AB_2564641 (BioLegend Cat. No. 801601)

Antigen Details

Structure
Neurofilament H has an apparent molecular mass of 200-220 kD.
Distribution

Tissue distribution: CNS, peripheral nerves and glandular cells of the prostate.
Cellular distribution: Cytoskeleton, nucleus, cytosol, and mitochondrion.

Function
Neurofilaments are the major components of the neuronal cytoskeleton. They provide axonal support and regulate axon diameter.
Interaction
Cell bodies and dendrites are generally unstained. Other cells and tissues are unreactive except for peripheral axons.
Cell Type
Mature Neurons
Biology Area
Cell Biology, Neuroscience, Neuroscience Cell Markers
Molecular Family
Intermediate Filaments, Phospho-Proteins
Antigen References

1. Petzold A. 2005. J. Neurol. Sci. 233 (1-2):183. PubMed

Gene ID
4744 View all products for this Gene ID
UniProt
View information about Neurofilament H on UniProt.org
Go To Top Version: 4    Revision Date: 01/17/2018

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This data display is provided for general comparisons between formats.
Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors.
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