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Immunofluorescence analysis of Raptor was performed using 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with Raptor Rabbit Polyclonal Antibody (424000) at 2ug/ml in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (A27034) 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 punctate cytoplasmic localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Mouse , Human , Rat|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic peptide derived from the C-terminal region of the human, mouse, and predicted rat raptor proteins|
|Storage buffer||PBS, pH 7.4|
|Contains||0.1% sodium azide|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||3-5 µg x 10^6 cells|
|Immunocytochemistry (ICC)||2 µg/ml|
|Immunofluorescence (IF)||2 µg/ml|
|Immunoprecipitation (IP)||Assay Dependent|
|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.
Raptor is a binding partner of target of rapamycin (TOR). It binds to 4EBP1 and p70alpha. The binding of raptor to mTOR is essential for the mTOR-catalyzed phosphorylation of 4EBP1 in vitro. It enhances the mTOR kinase activity toward p70alpha. It is also an essential scaffold for the mTOR-catalyzed phosphorylation of 4EBP1 and mediates TOR action in vivo.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
RGD1311784, 4932417H02Rik, Mip1, Rap, Raptor, mKIAA1303, KOG1
p150 target of rapamycin (TOR)-scaffold protein containing WD-repeats, raptor, regulatory-associated protein of mTOR, KOG1, Mip1