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Human and Mouse Phospho-mTOR (Ser2448) and Total mTOR ELISA

Human and Mouse Phospho-mTOR (Ser2448) and Total mTOR ELISA

Hersteller	Raybiotech
Kategorie	Oncology By Cell Type Carcinoma Glioma Hematology Leukemia Lymphoma Myeloma Melanoma Cell Biology Cell Status ELISA Autophagy ELISA & Immunoassays ELISA Kits Immunoassay
Typ	Elisa-Kit
Specific against	Human, Mouse
Menge	2 Plate Kit
Host	Hamster - Armenian
ArtNr	PEL-mTOR-S2448-T-2
Targets	MTOR
eClass 6.1	32160605
eClass 9.0	32160605
Lieferbar
Species Detected	Human Mouse
Compatible Sample Types	Cell Lysates Tissue Lysates
Design Principle	Sandwich-based
Method of Detection	Colorimetric
Quantitative/Semi-Quantitative	Semi-Quantitative
Solid Support	96-well Microplate
Gene Symbols	FRAPFRAP1FRAP2MTORRAFT1RAPT1
Kit Components	Pre- Coated 96-well Strip Microplate Wash Buffer Anti- Phospho Antibody H R P- Conjugated Secondary Antibody Assay Diluent T M B One- Step Substrate Stop Solution Lysis Buffer Positive Control Sample
Other Materials Required	Distilled or deionized water100 ml and 1 liter graduated cylinders Tubes to prepare sample dilutions Protease and Phosphatase inhibitors Precision pipettes to deliver 2 µl to 1 ml volumes Adjustable 1-25 ml pipettes for reagent preparation Benchtop rocker or shaker Microplate reader capable of measuring absorbance at 450 nm
Protocol Outline	Prepare all reagents and samples as instructed in the manual. Add 100 µl of sample or positive control to each well. Incubate 2.5 h at R T or O/ N at 4 ° C. Add 100 µl of prepared primary antibody to each well. Incubate 1 h at R T. Add 100 µl of prepared 1 X H R P- Streptavidin to each well. Incubate 1 h at R T. Add 100 µl of T M B One- Step Substrate Reagent to each well. Incubate 30 min at R T. Add 50 µl of Stop Solution to each well. Read at 450 nm immediately.
Protein Name & Synonyms	Serine/threonine-protein kinase mTOR (EC 2.7.11.1) (FK506-binding protein 12-rapamycin complex-associated protein 1) (FKBP12-rapamycin complex-associated protein) (Mammalian target of rapamycin) (mTOR) (Mechanistic target of rapamycin) (Rapamycin and FKBP12 target 1) (Rapamycin target protein 1)
Pathway	AMPK SignalingAutophagyPI3K-AKT Signaling
Positive Control	A431 cells were treated with recombinant human EGF at 37°C for 20 min. Cells were solubilzed at 4 x 107 cells/ml in Cell Lysate Buffer. Serial dilutions of lysates were analyzed in this ELISA (see Reagent Preparation step 4).
FBS Stimulation of 293 Cell Lines	293 cells were untreated or treated with 20% FBS for 20 min. Cell lysates were analyzed using this phosphoELISA. A431 cells were treated with recombinant human EGF at 37°C for 20 min. Cell lysates were analyzed using Western Blot.

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