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Immunofluorescence analysis of Neurofilament, Light chain was done on 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 Neurofilament, Light chain (DA2) Mouse Monoclonal Antibody (MA12010) at 2ug/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 (Product # A28175) at 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 is a merged image showing cytoplasmic localization. Panel e is a no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Human, Mouse, Rat|
|Published species reactivity||Chicken|
|Host / Isotype||Mouse|
|Immunogen||Enzymatically dephosphorylated porcine neurofilaments.|
|Storage buffer||PBS with 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|ELISA (ELISA)||Assay dependent|
|Flow Cytometry (Flow)||3-5 µg/10^6 cells|
|Immunocytochemistry (ICC)||2-3 µg/mL|
|Immunofluorescence (IF)||2-3 µg/mL|
|Immunohistochemistry (Frozen) (IHC (F))||Assay Dependent|
|Immunohistochemistry (Paraffin) (IHC (P))||Assay Dependent|
|Western Blot (WB)||1-2 µg/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.
|Immunocytochemistry (ICC)||See 1 publications below|
MA1-2010 detects the neurofilament, light chain in human, rat, and mouse samples.
MA1-2010 has been successfully used in western blot, immunohistochemistry, and immunofluorescence. By western blot this antibody detects a ~70 kDa protein representing the neurofilament, light chain in mouse brain tissue lysate. In immunohistochemistry procedures MA1-2010 recognizes the neurofilament, light chain in mouse brain tissue. In immunofluorescence procedures MA1-2010 recognizes the neurofilament, light chain in rat cerebral cortices.
The MA1-2010 immunogen is enzymatically dephosphorylated porcine neurofilaments.
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-L is the light or low molecular weight microfilament subunit and runs on SDS-PAGE gels at approximately 70 kDa.
Like most other intermediate filament proteins (IFPs), the expression of the different neuronal IFPs is both tissue-specific and developmentally regulated. The neurofilament (NF) triplet proteins (~70, 160, and 200 kDa) occur in both the central and peripheral nervous system and are normally restricted to neurons. The 70 kDa NF-protein can self-assemble into a filamentous structure, whereas the 160 kDa and 200 kDa NF-proteins require the presence of the 70 kDa NF-protein to co-assemble.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Identification of a tektin-like protein associated with neurofilaments in the developing chick nervous system.
MA1-2010 was used in immunocytochemistry to investigate the association between tektin-like protein and neurofilaments in chicken neurons
|Edson KJ,Linck RW,Letourneau PC||Journal of neuroscience research (30:105)||1991|
68 kDa neurofilament protein; 70 kDa neurofilament protein; CMT1F; CMT2E; light molecular weight neurofilament protein; neurofilament light polypeptide; neurofilament protein L; neurofilament protein, light chain; neurofilament subunit NF-L; Neurofilament triplet L pro; neurofilament triplet L protein; neurofilament, light polypeptide 68kDa; NFL; NLC; protein phosphatase 1, regulatory subunit 110
AI847934; CMT1F; CMT2E; NEFL; NF-L; NF68; NFL; PPP1R110