Immunofluorescence analysis of mTOR was done on 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 15 minutes, permeabilized with 0.25% Triton™ X-100 for 10 minutes, and blocked with 5% BSA for 1 hour at room temperature. The cells were labeled with mTOR Mouse monoclonal Antibody (AHO1232) at 2µg/mL in 1% BSA and incubated for 3 hours at room temperature and then labeled with Alexa Flour 488 Rabbit Anti-Mouse IgG Secondary Antibody (A11059) at a dilution of 1:400 for 30 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant DAPI (S36938). F-actin (Panel c: red) was stained with Alexa Fluor 594 Phalloidin (A12381). Panel d is a merged image showing cytoplasmic localization. Panel e shows no primary antibody control. The images were captured at 20X magnification.
|Tested species reactivity||Human, Mouse, Rat|
|Published species reactivity||Rat|
|Host / Isotype||Mouse / IgG2b, kappa|
|Immunogen||Recombinant fragment of human mTOR expressed in E. coli.|
|Storage buffer||PBS, pH 7.2, with 1% BSA|
|Contains||0.1% sodium azide|
|Tested Applications||Dilution *|
|Immunohistochemistry (Paraffin) (IHC (P))||1:10-1:100|
|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.
|Western Blot (WB)||See 1 publications below|
Mammalian target of rapamycin (mTOR) is a serine/threonine kinase that plays a key role in cell growth, cell proliferation, and protein synthesis. mTOR mediates phosphoinositide 3-kinase and Akt/PKB signaling, resulting in phosphorylation of 4EBP1, and initiation of mRNA translation. A second pathway involves regulation of ribosomal S6 kinase, which affects ribosome biogenesis and translation elongation.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Rapamycin reveals an mTOR-independent repression of Kv1.1 expression during epileptogenesis.
AHO1232 was used in western blot to study mTOR, microRNA miR-129-5p, and Kv1.1 in epilepsy
|Sosanya NM,Brager DH,Wolfe S,Niere F,Raab-Graham KF||Neurobiology of disease (73:96)||2015|