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Immunofluorescence analysis of alpha-Internexin was performed using 70% confluent log phase SH-SY5Y cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with alpha-Internexin (2E3) Mouse Monoclonal Antibody (32-3600) at 2 µg/ml in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (A28175) a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Human, Mouse|
|Published species reactivity||Human, Mouse|
|Host / Isotype||Mouse / IgG1|
|Immunogen||Full-length recombinant rat alpha-internexin protein.|
|Storage buffer||PBS, pH 7.4|
|Contains||0.1% sodium azide|
|Tested Applications||Dilution *|
|ELISA (ELISA)||1.0 ug/ml|
|Flow Cytometry (Flow)||3-5 µg/10^6 cells|
|Immunocytochemistry (ICC)||2 µg/ml|
|Immunofluorescence (IF)||2 µg/ml|
|Immunohistochemistry (IHC)||Assay Dependent|
|Western Blot (WB)||0.1-0.5 ug/ml|
* 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.
Alpha internexin is a Class IV intermediate filament originally discovered as it copurifies with other neurofilament subunits. On SDS-PAGE gels it runs with an apparent molecular weight of 64 to 66 kDa, with some species variability, although the real molecular weight is about 55kDa; as with the other neurofilament subunits the presence of highly negatively charged sequences results in reduction of SDS-PAGE mobility. a-internexin is related to but distinct from the better known neurofilament triplet proteins, NF-L, NF-M and NF-H, having similar protein sequence motifs and a similar intron organization. It is expressed only in neurons and in large amounts early in neuronal development, but is down-regulated in many neurons as development proceeds. Many classes of mature neurons contain a-internexin in addition to NF-L, NF-M and NF-H. In some mature neurons a-internexin is the only neurofilament subunit expressed. Antibodies to a-internexin are therefore unique probes to study and classify neuronal types and follow their processes in sections and in tissue culture. In addition, the very early developmental expression of a-internexin means its presence is an early and convenient diagnostic feature of neuronal progenitors cells and other cells committed to the neuronal lineage. In addition, recent studies show a marked up-regulation of a-internexin during neuronal regeneration.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
An autopsy case of neuronal intermediate filament inclusion disease with regard to immunophenotypic and topographical analysis of the neuronal inclusions.
32-3600 was used in immunohistochemistry - paraffin section to report an autopsy case of neuronal intermediate filament inclusion disease.
|Inoue K,Fujimura H,Ueda K,Matsumura T,Itoh K,Sakoda S||Neuropathology : official journal of the Japanese Society of Neuropathology (35:545)||2015|
An MND/ALS phenotype associated with C9orf72 repeat expansion: abundant p62-positive, TDP-43-negative inclusions in cerebral cortex, hippocampus and cerebellum but without associated cognitive decline.
32-3600 was used in immunohistochemistry - paraffin section to study 4 cases of chromosome 9-linked motor neuron disease/amyotrophic lateral sclerosis.
|Troakes C,Maekawa S,Wijesekera L,Rogelj B,Siklós L,Bell C,Smith B,Newhouse S,Vance C,Johnson L,Hortobágyi T,Shatunov A,Al-Chalabi A,Leigh N,Shaw CE,King A,Al-Sarraj S||Neuropathology : official journal of the Japanese Society of Neuropathology (32:505)||2012|
PRKAR1B mutation associated with a new neurodegenerative disorder with unique pathology.
32-3600 was used in western blot to study the association of PRKAR1B mutation with a new neurodegenerative disorder
|Wong TH,Chiu WZ,Breedveld GJ,Li KW,Verkerk AJ,Hondius D,Hukema RK,Seelaar H,Frick P,Severijnen LA,Lammers GJ,Lebbink JH,van Duinen SG,Kamphorst W,Rozemuller AJ,Bakker EB,Neumann M,Willemsen R,Bonifati V,Smit AB,van Swieten J||Brain : a journal of neurology (137:1361)||2014|
|Human||Not Cited||FET proteins TAF15 and EWS are selective markers that distinguish FTLD with FUS pathology from amyotrophic lateral sclerosis with FUS mutations.||Neumann M,Bentmann E,Dormann D,Jawaid A,DeJesus-Hernandez M,Ansorge O,Roeber S,Kretzschmar HA,Munoz DG,Kusaka H,Yokota O,Ang LC,Bilbao J,Rademakers R,Haass C,Mackenzie IR||Brain : a journal of neurology (134:2595)||2011|
Gliosarcoma with ependymal and PNET-like differentiation.
32-3600 was used in immunohistochemistry to characterize a rare case of gliosarcoma
|Shintaku M,Yoneda H,Hirato J,Nagaishi M,Okabe H||Clinical neuropathology (32:508)||2013|
||FET proteins TAF15 and EWS are selective markers that distinguish FTLD with FUS pathology from amyotrophic lateral sclerosis with FUS mutations.||Neumann M,Bentmann E,Dormann D,Jawaid A,DeJesus-Hernandez M,Ansorge O,Roeber S,Kretzschmar HA,Munoz DG,Kusaka H,Yokota O,Ang LC,Bilbao J,Rademakers R,Haass C,Mackenzie IR||Brain : a journal of neurology (134:2595)||2011|
Cytoskeletal organization of the developing mouse olfactory nerve layer.
32-3600 was used in immunohistochemistry to test if the cytoskeletal elements of olfactory sensory neuron axons may be are differentially expressed across the olfactory nerve layer.
|Akins MR,Greer CA||The Journal of comparative neurology (494:358)||2006|
|Human||Not Cited||What's in a name? Neuronal intermediate filament inclusion disease (NIFID), frontotemporal lobar degeneration-intermediate filament (FTLD-IF) or frontotemporal lobar degeneration-fused in sarcoma (FTLD-FUS)?||Menon R,Baborie A,Jaros E,Mann DM,Ray PS,Larner AJ||Journal of neurology, neurosurgery, and psychiatry (82:1412)||2011|
66 kDa neurofilament protein; Alpha-internexin; Alpha-Inx; internexin neuronal intermediate filament protein, alpha; NEF5; Neurofilament 5; neurofilament 5 (66kD); Neurofilament-66; neurofilament-66, tax-binding protein; NF-66
AV028420; INA; NEF5; NF-66; NF66; TXBP-1