Immunofluorescent analysis of the neurofilament medium chain in paraffin-embedded human brain tissue (right) compared to a negative control without primary antibody (left). Tissue sections were deparaffinized with xylene, and rehydrated with ethanol. To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0) and microwaved for 8-15 min. Following antigen retrieval, tissues were washed with water and PBS, and then blocked in 0.3% BSA for 30 min at room temperature. Tissues were then probed with a neurofilament medium chain monoclonal antibody (Product # 13-0500) in 0.3% BSA at a dilution of 1:50 for 1 hour at 37°C. Tissues were then incubated with a Goat anti-Mouse IgG (H+L) Secondary Antibody, DyLight 488 conjugate for 1 hour at 37°C (green). Nuclei (blue) were stained with DAPI. Images were taken at 40X magnification.
|Tested species reactivity||Human, Leech, Rat|
|Published species reactivity||Rat, Zebrafish, Mouse, Human, Not Applicable|
|Host / Isotype||Mouse / IgG1, kappa|
|Contains||0.1% sodium azide|
|Tested Applications||Dilution *|
|Immunohistochemistry (IHC)||5-10 ug/ml|
|Immunoprecipitation (IP)||2-5 ug|
|Western Blot (WB)||Assay Dependent|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
Involved in the maintenance of neuronal caliber, neurofilaments are the intermediate filament proteins found specifically in neurons, and are composed predominantly of three major proteins called NF-L, NF-M and NF-H. Like most other intermediate filament proteins (IFPs), the expression of the different neuronal IFPs is both tissue-specific and developmentally regulated. NF-M is the medium molecular weight microfilament subunit and runs on SDS-PAGE gels at approximately 160 kDa.
Neurofilament are the 10nm or intermediate filament proteins found specifically in neurons, and are composed predominantly of three major proteins called NF-L, NF-M and NF-H. NF-H is the heavy or high molecular weight microfilament subunit and runs on SDS-PAGE gels in the range 180-220kDa, with some variation in different species. NF-H polyclonal antibody can be used to identify neurons and their processes in tissue sections and in tissue culture and the antibody can also be used to study microfilament accumulations seen in many neurological diseases, such as Lou Geri's disease or Alzheimer's disease.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Rapid reverse genetic screening using CRISPR in zebrafish.
13-0500 was used in immunohistochemistry to identify novel genes involved in electrical-synapse formation using CRISPR in zebrafish.
|Shah AN,Davey CF,Whitebirch AC,Miller AC,Moens CB||Nature methods (12:535)||2015|
Unique function of Kinesin Kif5A in localization of mitochondria in axons.
13-0500 was used in immunohistochemistry to characterize neuronal defects in zebrafish kif5Aa mutants
|Campbell PD,Shen K,Sapio MR,Glenn TD,Talbot WS,Marlow FL||The Journal of neuroscience : the official journal of the Society for Neuroscience (34:14717)||2014|
Characterization and classification of zebrafish brain morphology mutants.
13-0500 was used in immunohistochemistry to use zebrafish to identify genes required for brain morphogenesis
|Lowery LA,De Rienzo G,Gutzman JH,Sive H||Anatomical record (Hoboken, N.J. : 2007) (292:94)||2009|
Whitesnake/sfpq is required for cell survival and neuronal development in the zebrafish.
13-0500 was used in immunohistochemistry to use zebrafish to identify genes that contribute to brain organogenesis
|Lowery LA,Rubin J,Sive H||Developmental dynamics : an official publication of the American Association of Anatomists (236:1347)||2007|
|Zebrafish||Not Cited||The zebrafish Iroquois gene iro7 positions the r4/r5 boundary and controls neurogenesis in the rostral hindbrain.||Lecaudey V,Anselme I,Rosa F,Schneider-Maunoury S||Development (Cambridge, England) (131:3121)||2004|
vhnf1 and Fgf signals synergize to specify rhombomere identity in the zebrafish hindbrain.
13-0500 was used in immunohistochemistry to elucidate the mechanism of rhombomere determination.
|Wiellette EL,Sive H||Development (Cambridge, England) (130:3821)||2003|
|Not Applicable||Not Cited||
The C-terminal domains of NF-H and NF-M subunits maintain axonal neurofilament content by blocking turnover of the stationary neurofilament network.
13-0500 was used in western blot to determine the functions of heavy and medium neurofilament subunits
|Rao MV,Yuan A,Campbell J,Kumar A,Nixon RA||PloS one (7:null)||2012|
|Mouse||1:1000||Abnormal neurofilament transport caused by targeted disruption of neuronal kinesin heavy chain KIF5A.||Xia CH,Roberts EA,Her LS,Liu X,Williams DS,Cleveland DW,Goldstein LS||The Journal of cell biology (161:55)||2003|
Effect of nicotine and cocaine on neurofilaments and receptors in whole brain tissue and synaptoneurosome preparations.
13-0500 was used in western blot to assess the effect of repeated nicotine and cocaine administration on the expression of neurofilament proteins, actin, and alpha-7 nicotinic, dopamine D1, and NMDA NR1 receptors in brain
|Kovacs K,Lajtha A,Sershen H||Brain research bulletin (82:109)||2010|
Ammonium-induced impairment of axonal growth is prevented through glial creatine.
13-0500 was used in immunohistochemistry (frozen) to assess the effects of hyperammonemia in neonatal and infant rats.
|Braissant O,Henry H,Villard AM,Zurich MG,Loup M,Eilers B,Parlascino G,Matter E,Boulat O,Honegger P,Bachmann C||The Journal of neuroscience : the official journal of the Society for Neuroscience (22:9810)||2002|
Numerous conglomerate inclusions in slowly progressive familial amyotrophic lateral sclerosis with posterior column involvement.
13-0500 was used in immunohistochemistry (paraffin) to characterize disease in a patient with familial amyotrophic lateral sclerosis.
|Katayama S,Watanabe C,Noda K,Ohishi H,Yamamura Y,Nishisaka T,Inai K,Asayama K,Murayama S,Nakamura S||Journal of the neurological sciences (171:72)||1999|
Activation of a neurofilament kinase, a tau kinase, and a tau phosphatase by decreased ATP levels in nerve growth factor-differentiated PC-12 cells.
13-0500 was used in western blot to study the effects of a mitochondrial uncoupler on an Alzheimer's disease model.
|Bush ML,Miyashiro JS,Ingram VM||Proceedings of the National Academy of Sciences of the United States of America (92:1861)||1995|
|Not Applicable||Not Cited||
Ammonium alters creatine transport and synthesis in a 3D culture of developing brain cells, resulting in secondary cerebral creatine deficiency.
13-0500 was used in immunohistochemistry - frozen section to study rat brain cells treated with NH4+
|Braissant O,Cagnon L,Monnet-Tschudi F,Speer O,Wallimann T,Honegger P,Henry H||The European journal of neuroscience (27:1673)||2008|
|Zebrafish||1:25||FGF3 and FGF8 mediate a rhombomere 4 signaling activity in the zebrafish hindbrain.||Maves L,Jackman W,Kimmel CB||Development (Cambridge, England) (129:3825)||2002|