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Anti-Phospho-PRAS40 (Thr246) Rabbit Antibody [P11G2]

Catalog No.: F4154

Application: Reactivity: Human, Mouse, Rat, Monkey

Usage Information

Dilution
WB1:1000 IP1:50 IHC1:200

Application
WB, IP, IHC

Reactivity
Human, Mouse, Rat, Monkey

Source
Rabbit

Storage Buffer
PBS, pH 7.2+50% Glycerol+0.05% BSA+0.01% NaN3

Storage (from the date of receipt)
-20°C (avoid freeze-thaw cycles), 2 years

Predicted MW
40 kDa

Positive Control	Human breast carcinoma; Metastatic SK-OV-3 tumor; LNCaP cell; MCF-7 cell; HeLa cell (Insulin, 1μM, 30 min)
Negative Control	LNCaP cells (treated with LY294002)

Datasheet & SDS

Biological Description

Specificity
Anti-Phospho-PRAS40 (Thr246) Rabbit Antibody [P11G2] recognizes endogenous levels of PRAS40 protein only when phosphorylated at Thr246.

Clone
P11G2

Synonym(s)
Proline-rich AKT1 substrate 1, 40 kDa proline-rich AKT substrate, AKT1S1, PRAS40, 40 kDa AKT substrate, 40 kDa AKT substrate 1, AKT substrate 1

Background
PRAS40 (proline-rich AKT substrate of 40 kDa, encoded by AKT1S1) functions as a negative regulator of the mammalian target of rapamycin complex 1 (mTORC1). It exerts this effect by binding to Raptor and competing with mTOR substrates such as 4E-BP1 and p70S6K. Both PKB/AKT and mTOR phosphorylate PRAS40 at distinct residues—Thr246 by AKT, and Ser183/212/221 by mTOR—in vitro and in vivo. Phosphorylation of PRAS40 following AKT and mTOR activation promotes its dissociation from Raptor, thereby lifting its inhibitory effect and enabling mTORC1-driven phosphorylation of downstream effectors including 4E-BP1 and p70S6K. PRAS40 acts as an anti-apoptotic regulator during the early stages of embryoid body formation, which involves three-dimensional aggregates of pluripotent stem cells. In primary human skeletal muscle cells, PRAS40 downregulation reduces AKT phosphorylation as well as the activity of mTORC1-dependent targets. In cancer models, silencing PRAS40 has been shown to inhibit tumor growth in malignant melanoma and Ewing sarcoma. Phosphorylated PRAS40 also interacts with the transcription factor FOXO3a, suppressing transcription of pro-apoptotic genes. Elevated levels of phosphorylated PRAS40 have been linked to progression in several cancers, including melanoma, prostate, and gastric tumors. Conversely, expression of phosphorylation-deficient PRAS40 mutants blocks carcinogenesis. Importantly, phosphorylation of PRAS40 by AKT at Thr246 alleviates its repression of mTORC1, which in turn phosphorylates PRAS40 at Ser183, reinforcing the regulatory feedback.

Background

PRAS40 (proline-rich AKT substrate of 40 kDa, encoded by AKT1S1) functions as a negative regulator of the mammalian target of rapamycin complex 1 (mTORC1). It exerts this effect by binding to Raptor and competing with mTOR substrates such as 4E-BP1 and p70S6K. Both PKB/AKT and mTOR phosphorylate PRAS40 at distinct residues—Thr246 by AKT, and Ser183/212/221 by mTOR—in vitro and in vivo. Phosphorylation of PRAS40 following AKT and mTOR activation promotes its dissociation from Raptor, thereby lifting its inhibitory effect and enabling mTORC1-driven phosphorylation of downstream effectors including 4E-BP1 and p70S6K. PRAS40 acts as an anti-apoptotic regulator during the early stages of embryoid body formation, which involves three-dimensional aggregates of pluripotent stem cells. In primary human skeletal muscle cells, PRAS40 downregulation reduces AKT phosphorylation as well as the activity of mTORC1-dependent targets. In cancer models, silencing PRAS40 has been shown to inhibit tumor growth in malignant melanoma and Ewing sarcoma. Phosphorylated PRAS40 also interacts with the transcription factor FOXO3a, suppressing transcription of pro-apoptotic genes. Elevated levels of phosphorylated PRAS40 have been linked to progression in several cancers, including melanoma, prostate, and gastric tumors. Conversely, expression of phosphorylation-deficient PRAS40 mutants blocks carcinogenesis. Importantly, phosphorylation of PRAS40 by AKT at Thr246 alleviates its repression of mTORC1, which in turn phosphorylates PRAS40 at Ser183, reinforcing the regulatory feedback.

References
https://pubmed.ncbi.nlm.nih.gov/28945219/ https://pubmed.ncbi.nlm.nih.gov/32467173/

Reference Table for Selecting PVDF Membrane Pore Size Specifications

Protein Size (kDa)	PVDF Transfer Membrane Pore Size (μm)
< 10	0.22
10-20	0.22
20-30	0.45
30-45	0.45
45-70	0.45
80-100	0.45
100-140	0.45
> 140	0.45

Tech Support

Answers to questions you may have can be found in the inhibitor handling instructions. Topics include how to prepare stock solutions, how to store inhibitors, and issues that need special attention for cell-based assays and animal experiments.

Handling Instructions

Tel: +1-832-582-8158 Ext:3
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Protein Size (kDa)	Separating Gel Percentage
4-40	20%
12-45	15%
10-70	12.5%
15-100	10%
25-100	8%
60-210	5%