Key Product Details

Species Reactivity

Validated:

Human

Cited:

Human

Applications

Validated:

Western Blot, Immunocytochemistry, Simple Western

Cited:

Western Blot

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG1 Clone # 660928
View all PRAS40 Primary Antibodies »

Product Specifications

Immunogen

E. coli-dervied recombinant human PRAS40
Gln119-Tyr256
Accession # Q96B36

Specificity

Detects human PRAS40 in Western blots.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG1

Scientific Data Images for Human PRAS40 Antibody

Detection of Human PRAS40 antibody by Western Blot.

Detection of Human PRAS40 by Western Blot.

Western blot shows lysates of 293T human embryonic kidney cell line, A549 human lung carcinoma cell line, MDA-MB-468 human breast cancer cell line, and T47D human breast cancer cell line. PVDF Membrane was probed with 1 µg/mL of Mouse Anti-Human PRAS40 Monoclonal Antibody (Catalog # MAB6408) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for PRAS40 at approximately 40 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Detection of Human PRAS40 antibody by Simple WesternTM.

Detection of Human PRAS40 by Simple WesternTM.

Simple Western lane view shows lysates of 293T human embryonic kidney cell line, loaded at 0.5 mg/mL. A specific band was detected for PRAS40 at approximately 65 kDa (as indicated) using 10 µg/mL of Mouse Anti-Human PRAS40 Monoclonal Antibody (Catalog # MAB6408). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.
PRAS40 antibody in HeLa Human Cell Line by Immunocytochemistry (ICC).

PRAS40 in HeLa Human Cell Line.

PRAS40 was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line using Mouse Anti-Human PRAS40 Monoclonal Antibody (Catalog # MAB6408) at 8 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.

Applications for Human PRAS40 Antibody

Application
Recommended Usage

Immunocytochemistry

5-25 µg/mL
Sample: Immersion fixed HeLa human cervical epithelial carcinoma cell line

Simple Western

10 µg/mL
Sample: 293T human embryonic kidney cell line

Western Blot

1 µg/mL
Sample: 293T human embryonic kidney cell line, A549 human lung carcinoma cell line, MDA‑MB‑468 human breast cancer cell line, and T47D human breast cancer cell line

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. 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

UniProt

Q96B36 (Human)

Additional PRAS40 Products

Product Documents for Human PRAS40 Antibody

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Human PRAS40 Antibody

For research use only

Related Research Areas

Autophagy
Interferons

Citations for Human PRAS40 Antibody

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Protocols

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