Immunofluorescent analysis of mTOR (green) in HeLa cells. The cells were fixed with 4% paraformaldehyde, permeabilized with 0.1% Triton X-100 in PBS, and blocked with 1% Blocker BSA (Product # 37525), each for 15 minutes at room temperature. Cells were stained with an mTOR polyclonal antibody (Product # PA1-518) at a concentration of 10ug/ml in 1% Blocker BSA in PBS (right panel), or incubated in blocking buffer alone as a negative control (left panel) overnight at 4C, and then incubated with a Dylight 488-conjugated goat anti-rabbit IgG secondary antibody (Product # 35552) at a dilution of 1:1000 for 1 hour at room temperature. F-Actin (red) was stained with a DyLight 554-conjugated phalloidin control (Product # 21834) and nuclei (blue) were stained with DAPI (Product # 46190).
|Tested species reactivity||Human, Mouse, Non-human primate, Rat|
|Published species reactivity||Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Immunogen corresponds to amino acids 1223-1290 of human mTOR.|
|Storage buffer||PBS with 1mg/ml BSA, 30% glycerol|
|Contains||0.05% sodium azide|
|Tested Applications||Dilution *|
|Immunoprecipitation (IP)||2 ug|
|Western Blot (WB)||1:100-1:1000|
* 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|
3-8% Tris-Acetate Gel is recommended for Western blot.
PA1-518 detects a predominant band at ~290kD corresponding to mTOR. Other lower MW nonspecific bands of unknown identity were also detected at ~65kD and ~35kD in some cell lysates.
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.
IP-MS enrichment of MTOR (LFQ intensity): MTOR was enriched 74-fold from HCT116 lysate compared to background proteins, using the optimized IP-MS workflow with Pierce MS-Compatible Magnetic IP Kit protein A/G (Part No. 90409) and MTOR antibody (Part No. PA1-518). See more information on IP-MS verification of antibody selectivity. IP-MS validation info.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Bilirubin activates transcription of HIF-1¿ in human proximal tubular cells cultured in the physiologic oxygen content.
PA1-518 was used in western blot to investigate the effect of bilirubin on HIF-1 expression in proximal tubular cells cultured in physiologically relevant oxygen concentration
|Kim SG,Ahn SY,Lee ES,Kim S,Na KY,Chae DW,Chin HJ||Journal of Korean medical science (29 Suppl 2:S146)||2014|