Human/Mouse RPTOR Antibody

Catalog #: MAB5957 Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
MAB5957
MAB5957-SP

Detection of Human and Mouse RPTOR by Western Blot.

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Summary Data Examples Preparation and Storage Reconstitution Calculator Background Product Datasheets Related Research Areas

Human/Mouse RPTOR Antibody Summary

Species Reactivity

Human, Mouse

Specificity

Detects human and mouse RPTOR in Western blots.

Source

Monoclonal Mouse IgG_2B Clone # 514208

Purification

Protein A or G purified from hybridoma culture supernatant

Immunogen

E. coli-derived recombinant human RPTOR
Lys77-Gln230
Accession # Q8N122

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Label

Unconjugated

Applications

Recommended Concentration

Sample

Western Blot

1 µg/mL

See below

CyTOF-ready

Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

Intracellular Staining by Flow Cytometry

0.25 µg/10⁶ cells

See below

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Western Blot

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Detection of Human and Mouse RPTOR by Western Blot. Western blot shows lysates of 293T human embryonic kidney cell line, HeLa human cervical epithelial carcinoma cell line, and Neuro-2A mouse neuroblastoma cell line. PVDF Membrane was probed with 1 µg/mL of Human/Mouse RPTOR Monoclonal Antibody (Catalog # MAB5957) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for RPTOR at approximately 150 kDa (as indicated). This experiment was conducted under non-reducing conditions and using Immunoblot Buffer Group 1.

Intracellular Staining by Flow Cytometry

Detection of RPTOR antibody in HeLa Human Cell Line antibody by Flow Cytometry.

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Detection of RPTOR in HeLa Human Cell Line by Flow Cytometry. HeLa human cervical epithelial carcinoma cell line was stained with Mouse Anti-Human/Mouse RPTOR Monoclonal Antibody (Catalog # MAB5957, filled histogram) or isotype control antibody (Catalog # MAB0041, open histogram), followed by Phycoerythrin-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0102B). To facilitate intracellular staining, cells were fixed with Flow Cytometry Fixation Buffer (Catalog # FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (Catalog # FC005). View our protocol for Staining Intracellular Molecules.

Intracellular Staining by Flow Cytometry

Detection of RPTOR antibody in C2C12 Mouse Cell Line antibody by Flow Cytometry.

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Detection of RPTOR in C2C12 Mouse Cell Line by Flow Cytometry. C2C12 mouse myoblast cell line was stained with Mouse Anti-Human/Mouse RPTOR Monoclonal Antibody (Catalog # MAB5957, filled histogram) or isotype control antibody (Catalog # MAB0041, open histogram), followed by Phycoerythrin-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0102B). To facilitate intracellular staining, cells were fixed with Flow Cytometry Fixation Buffer (Catalog # FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (Catalog # FC005). View our protocol for Staining Intracellular Molecules.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.

12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: RPTOR

RPTOR (Raptor) is a 150 kDa component of the cytosolic mammalian target of Rapamycin complex 1 (mTORC1) which also contains mTOR and GBL proteins. mTORC1 plays a dominant role in cell cycle regulation in response to metabolic conditions. The interaction of RPTOR with the kinase mTOR is stabilized under conditions of nutrient deprivation and energy stress, leading to inhibition of mTOR and cell cycle arrest. RPTOR contains multiple Ser and Thr residues whose phosphorylation regulates the activation status of mTOR. RPTOR is critical for the response of skeletal muscle and adipose tissue to insulin. It contains three RNC blocks (aa 48-511), three HEAT repeats (aa 550-667), and seven C-terminal WD40 domains (aa 1020-1335). Within aa 77-230, human RPTOR shares 100% aa sequence identity with mouse and rat RPTOR.

Long Name

Regulatory Associated Protein of mTOR

Entrez Gene IDs

57521 (Human); 74370 (Mouse); 287871 (Rat)

Alternate Names

KIAA1303; KIAA1303p150 target of rapamycin (TOR)-scaffold protein containing WD-repeats; KOG1; Mip1; p150 target of rapamycin (TOR)-scaffold protein; Raptor; regulatory associated protein of MTOR, complex 1; regulatory-associated protein of mTOR; RPTOR