Immunofluorescence analysis of Neurofibromin 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 Neurofibromin (McNFn27b) Mouse Monoclonal Antibody (MA1085) 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 punctate cytoplasmic localization. Panel e is a no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Human, Mouse, Rat|
|Host / Isotype||Mouse / IgG1|
|Immunogen||Synthetic peptide corresponding to the N-terminal residues 27-41 of human neurofibromin.|
|Storage buffer||PBS with 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Immunocytochemistry (ICC)||1-2 µg/ml|
|Immunofluorescence (IF)||1-2 µg/ml|
|Immunohistochemistry (IHC)||5-10 ug/mL|
|Western Blot (WB)||1-3 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.
MA1-085 detects neurofibromin in human, rat, and mouse cells.
MA1-085 has been successfully used in Western blot, and immunohistochemistry procedures. By Western blot, this antibody detects a ~240-260 kDa protein.
The MA1-085 immunogen is a synthetic peptide corresponding to the N-terminal residues 27-41 of human neurofibromin.
Neurofibromin is a member of the GTPase-activating (GAP) class of proteins. These proteins enhance the GTPase activity of signaling G-Proteins like Ras. The NF1 gene is also classified as a tumor suppressor gene, and many studies have shown that mutations at the NF1 locus produce neurofibromas, rhabdomyosarcomas, leimyosarcomas, mental retardation and memory deficiency. Neurofibromin has also been associated with the intermediated filaments of the cytoskeleton. NF1 is expressed at its highest levels during periods of desmosome formation and mutations of NF1 produce variable cell sizes and morphologies.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.