Rabbit anti-Human, Mouse PRAS40 Polyclonal Antibody | anti-AKT1S1 antibody

PRAS40, phosphorylated (Thr246) (Proline-Rich AKT Substrate 40kD Lobe, AKT151)

Cat. Num. AAA613629

• Gene Names: AKT1S1; Lobe; PRAS40
• Reactivity: Human, Mouse
• Applications: Western Blot
• Purity: Affinity Purified; Purified by immunoaffinity chromatography.
• Synonyms: PRAS40; Polyclonal Antibody; phosphorylated (Thr246) (Proline-Rich AKT Substrate 40kD Lobe; AKT151); Anti -PRAS40; anti-AKT1S1 antibody

• Host: Rabbit
• Reactivity: Human, Mouse
• Clonality: Polyclonal
• Specificity: Recognizes phosphorylated PRAS40 (Thr246)), Mr 40kDa. Species Cross-reactivity: Human and mouse tested.
• Purity/Purification: Affinity Purified; Purified by immunoaffinity chromatography.
• Form/Format: Supplied as a liquid in PBS (without Mg2+ and Ca2+), pH 7.3 containing 1.0 mg/mL BSA (IgG, protease free) and 0.05% sodium azide and 50% glycerol.

• Applicable Applications for anti-AKT1S1 antibody: Western Blot (WB)
• Application Notes: Suitable for use in Western Blot.; Dilution: Western Blot: 1:1000
• Immunogen: Peptide corresponding to amino acid region encompassing the human phospho-PRAS40 (Thr246).
• Preparation and Storage: May be stored at 4 degree C for short-term only. For long-term storage, store at -20 degree C. Aliquots are stable for at least 12 months at -20 degree C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.

Western Blot (WB)

(Western Blot Analysis Untreated (Lane 1) or PDGF treated (Lanes 2, 3, 4) lysates from NIH3T3 cells were resolved by electrophoresis, transferred to nitrocellulose, and probed with anti-phospho-PRAS40 (Thr246) for 2 hours at room temperature. Lane 3 was pre-incubated with the blocking peptide immunogen corresponding to phospho- PRAS40 (Thr246). Lane 4 blot was pre-incubated with λ phosphatase. Proteins were visualized via HRP and chemiluminescent detection. Arrow indicates phospho-PRAS40 (Thr246).)

Related Product Information for anti-AKT1S1 antibody

The proline-rich Akt substrate of 40 kilodaltons (PRAS40) was identified as a raptor binding protein that is phosphorylated directly by mTOR complex (mTORC) 1 but not mTORC 2 in vitro, predominantly at PRAS40 (Ser183). The binding of S6K1 and 4E-BP1 to raptor requires a "TOR signaling" (TOS) motif which contains an essential Phe followed by four alternating acidic and small hydrophobic amino acids; PRAS40 binding to raptor was severely inhibited by mutation of PRAS40 (Phe129) to Ala; immediately carboxyterminal to Phe129 are two small hydrophobic amino acid followed by two acidic residues, an arrangement different from that of the canonical TOS motif. PRAS40 binding to raptor was also abolished by mutation of the major mTORC1 phosphorylation site Ser183, to Asp. PRAS40 (Ser183) was phosphorylated in intact cells; as with S6K1 and 4E-BP1, PRAS40 (Ser183) phosphorylation was inhibited by rapamycin, by 2-deoxyglucose and by overexpression of the tuberous sclerosis complex heterodimer. PRAS40 (Ser183) phosphorylation was also inhibited reversibly by withdrawal of all or of only the branched-chain amino acids; this inhibition was reversed by overexpression of the Rheb GTPase. Overexpressed PRAS40 suppressed the phosphorylation of S6K1 and 4E-BP1 at their rapamycin-sensitive phosphorylation sites, and reciprocally, overexpression of S6K1 or 4E-BP1 suppressed phosphorylation of PRAS40 (Ser183) and its binding to raptor. More importantly, RNAi-induced depletion of PRAS40 enhanced the amino-acid stimulated phosphorylation of both S6K1 and 4E-BP1. These results establish PRAS40 as a physiological mTORC1 substrate that contains a variant TOS motif. Moreover, they indicate that the capacity or ability of raptor to bind endogenous substrates is limiting for the activity of mTORC1 in vivo, and is therefore a potential locus of regulation, as suggested by others.

Product Categories/Family for anti-AKT1S1 antibody

Antibodies; Abs to Protein Kinase Substrates

NCBI and Uniprot Product Information

• NCBI GI #: 498917186
• NCBI GeneID: 84335
• NCBI Accession #: NP_001265089.1
• NCBI GenBank Nucleotide #: NM_001278160.1
• UniProt Accession #: Q96B36; Q96BI4; Q96IK7; Q96NG2; Q9BWR5; A8MTQ1; B2RE93; J3KPM3
• Molecular Weight: 27,383 Da
• NCBI Official Full Name: proline-rich AKT1 substrate 1 isoform b
• NCBI Official Synonym Full Names: AKT1 substrate 1 (proline-rich)
• NCBI Official Symbol: AKT1S1
• NCBI Official Synonym Symbols: Lobe; PRAS40
• NCBI Protein Information: proline-rich AKT1 substrate 1; 40 kDa proline-rich AKT substrate
• UniProt Protein Name: Proline-rich AKT1 substrate 1
• Protein Family: Proline-rich AKT1 substrate
• UniProt Gene Name: AKT1S1
• UniProt Synonym Gene Names: PRAS40
• UniProt Entry Name: AKTS1_HUMAN

NCBI Description

AKT1S1 is a proline-rich substrate of AKT (MIM 164730) that binds 14-3-3 protein (see YWHAH, MIM 113508) when phosphorylated (Kovacina et al., 2003 [PubMed 12524439]).[supplied by OMIM, Mar 2008]

Uniprot Description

Function: Subunit of mTORC1, which regulates cell growth and survival in response to nutrient and hormonal signals. mTORC1 is activated in response to growth factors or amino acids. Growth factor-stimulated mTORC1 activation involves a AKT1-mediated phosphorylation of TSC1-TSC2, which leads to the activation of the RHEB GTPase that potently activates the protein kinase activity of mTORC1. Amino acid-signaling to mTORC1 requires its relocalization to the lysosomes mediated by the Ragulator complex and the Rag GTPases. Activated mTORC1 up-regulates protein synthesis by phosphorylating key regulators of mRNA translation and ribosome synthesis. mTORC1 phosphorylates EIF4EBP1 and releases it from inhibiting the elongation initiation factor 4E (eiF4E). mTORC1 phosphorylates and activates S6K1 at 'Thr-389', which then promotes protein synthesis by phosphorylating PDCD4 and targeting it for degradation. Within mTORC1, AKT1S1 negatively regulates mTOR activity in a manner that is dependent on its phosphorylation state and binding to 14-3-3 proteins. Inhibits RHEB-GTP-dependent mTORC1 activation. Substrate for AKT1 phosphorylation, but can also be activated by AKT1-independent mechanisms. May also play a role in nerve growth factor-mediated neuroprotection. Ref.7 Ref.9 Ref.10

Subunit structure: Part of the mammalian target of rapamycin complex 1 (mTORC1) which contains MTOR, MLST8, RPTOR, AKT1S1/PRAS40 and DEPTOR. mTORC1 binds to and is inhibited by FKBP12-rapamycin. Interacts directly with RPTOR. The phosphorylated form interacts with 14-3-3 proteins. Ref.6 Ref.9 Ref.10

Subcellular location: Cytoplasm › cytosol

By similarity. Note: Found in the cytosolic fraction of the brain

By similarity.

Tissue specificity: Widely expressed with highest levels of expression in liver and heart. Expressed at higher levels in cancer cell lines (e.g. A-549 and HeLa) than in normal cell lines (e.g. HEK293). Ref.6 Ref.7

Post-translational modification: Phosphorylated by AKT1. Phosphorylation relieves inhibitory function on mTORC1. Ref.6

Sequence caution: The sequence AAH00031.2 differs from that shown. Reason: Erroneous initiation.