Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Human, Mouse

Cited:

Human

Applications

Validated:

Western Blot, Immunocytochemistry

Cited:

Western Blot

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG2A Clone # 760502
View all PRAS40 Primary Antibodies »

Product Specifications

Immunogen

Phosphopeptide containing the human PRAS40 T246 site

Specificity

Detects human and mouse PRAS40 when phosphorylated at T246

Clonality

Monoclonal

Host

Mouse

Isotype

IgG2A

Scientific Data Images for Human/Mouse Phospho-PRAS40 (T246) Antibody

Detection of Human and Mouse Phospho-PRAS40 (T246) antibody by Western Blot.

Detection of Human and Mouse Phospho-PRAS40 (T246) by Western Blot.

Western blot shows lysates of MCF-7 human breast cancer cell line and NIH-3T3 mouse embryonic fibroblast cell line untreated (-) or treated (+) with 100 ng/mL Recombinant Human IGF-I (Catalog # 291-G1) for 20 minutes and 10 ng/mL Recombinant Human PDGF-BB (Catalog # 220-BB) for 5 minutes. PVDF membrane was probed with 0.5 µg/mL of Mouse Anti-Human Phospho-PRAS40 (T246) Monoclonal Antibody (Catalog # MAB6890) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for Phospho-PRAS40 (T246) at approximately 40 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Phospho-PRAS40 (T246) antibody in MCF-7 Human Cell Line by Immunocytochemistry (ICC).

Phospho-PRAS40 (T246) in MCF‑7 Human Cell Line.

PRAS40 phosphorylated at T246 was detected in immersion fixed MCF-7 human breast cancer cell line using Mouse Anti-Human/Mouse Phospho-PRAS40 (T246) Monoclonal Antibody (Catalog # MAB6890) at 25 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Detection of Human PRAS40 by Western Blot

Detection of Human PRAS40 by Western Blot

PIK3R2 depletion, but not PI3K inhibitors, induces stable PI3K pathway inhibitionA. Tumor xenografts were obtained using H226, CaLu-1 and H520 lung SQCC cells expressing inducible PIK3R1 or PIK3R2 shRNA. When tumors reached ~50 mm3, mice were treated with doxycycline in drinking water (2 mg/ml); they were sacrificed when tumors began to diminish. Normalized tumor extracts were examined by WB with indicated antibodies. B. H226, CaLu-1 and H520 cells were cultured in exponential growth were incubated with: vehicle (DMSO, 1:103 V:V), Ly294002 (5 μM), PIK75 (200 nM), TGX221 (30 μM), or PIK75 (200 nM) plus TGX221 (30 μM) for the last 1 h of culture or during 48 h; extracts were tested in WB. Mr indicates relative mobility. In both A and B, the pAkt or pPRAS40 signal was measured and normalized to the loading control (Akt or tubulin, respectively), and compared to that in controls (DMSO, considered 100%). Graphs show percentage of signal compared to maximal as mean ± SD at 48h of treatment. Arrows on the right side of bars show the percentage of signal compared to maximal detected after 1h of treatment.* P <0.05; **P <0.01; unpaired Student's t test with Welch correction. Image collected and cropped by CiteAb from the following publication (https://www.oncotarget.com/lookup/doi/10.18632/oncotarget.13195), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Human/Mouse Phospho-PRAS40 (T246) Antibody

Application
Recommended Usage

Immunocytochemistry

8-25 µg/mL
Sample: Immersion fixed MCF‑7 human breast cancer cell line

Western Blot

0.5 µg/mL
Sample: MCF‑7 human breast cancer cell line and NIH‑3T3 mouse embryonic fibroblast cell line treated with Recombinant Human IGF‑I (Catalog # 291-G1) and Recombinant Human PDGF‑BB (Catalog # 220-BB)

Reviewed Applications

Read 1 review rated 5 using MAB6890 in the following applications:

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Sterile PBS to a final concentration of 0.5 mg/mL. For liquid material, refer to CoA for concentration.

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.

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.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: PRAS40

PRAS40 (Proline-rich AKT1 substrate 1; also Akt1S1 and p39) is a 40-42 kDa cytoplasmic phosphoprotein that lacks generally recognized structural motifs. It is widely expressed and is considered to be key regulator of mTORC1 (mTOR plus raptor and G beta L), a complex through which Akt signals into the cell. Through phosphorylation, mTORC1 activity is upregulated by PRAS40. In particular, nonphosphorylated PRAS40 binds to and serves as a negative regulator of mTORC1 activity. Upon Insulin signaling, PRAS40 is phosphorylated on Thr246, Ser221 and Ser183. This causes it to bind 14-3-3 and results in its dissociation from mTORC1, freeing up mTOR to regulate (positively or negatively) protein synthesis. Human PRAS40 is 256 amino acids (aa) in length. It contains one extended Pro-rich region (aa 35-96) plus at least nine utilized Ser/Thr phosphorylation sites. There is one alternative start site at Met131. Over aa 119-256, human PRAS40 shares 93% aa identity with mouse PRAS40.

Long Name

40 kDa Proline-rich Akt1 Substrate

Alternate Names

AKT1S1, Lobe

Entrez Gene IDs

84335 (Human); 67605 (Mouse); 292887 (Rat)

Gene Symbol

AKT1S1

Additional PRAS40 Products

Product Documents for Human/Mouse Phospho-PRAS40 (T246) Antibody

Certificate of Analysis

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Product Specific Notices for Human/Mouse Phospho-PRAS40 (T246) Antibody

For research use only

Related Research Areas

Autophagy
Interferons

Citations for Human/Mouse Phospho-PRAS40 (T246) Antibody

Customer Reviews for Human/Mouse Phospho-PRAS40 (T246) Antibody (1)

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  • Human/Mouse Phospho-PRAS40 (T246) Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: MCF-7 human breast cancer cell line
    Species: Human
    Verified Customer | Posted 12/23/2021
    Human/Mouse Phospho-PRAS40 (T246) Antibody MAB6890

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