Invitrogen

Phospho-AS160 (Thr642) Polyclonal Antibody

Product Image Gallery

Phospho-AS160 (Thr642) Antibody — Fig 2 Phosphorylation of AS160/TBC1D4 in mouse heart in response to insulin. PAS-binding signals, phosphorylation of AS160-Thr 649 and PKB-Ser 473 , and total AS160 and PKB were detected in the lysates (40 mug) of hearts from the wild-type and AS160 Thr649Ala knockin mice that were intraperitoneally injected with PBS (basal) or insulin (150 mU per kg of body weight) for 20 min. GAPDH was used as a loading control.

Phospho-AS160 (Thr642) Antibody in Western Blot (WB) — Western Blotting. and Mutant Analysis Extracts of 3T3-L1 cells (1-4) or COS cells over-expressing WT AS160 (5, 6) or mutant AS160 T642A (7), treated with PI-3-Kinase inhibitor LY294002 (5) or insulin (1-4, 6-7), were resolved by SDS-PAGE on a 10% Tris-glycine gel and transferred to PVDF. The membrane was blocked with a 5% BSA-TBST buffer for one hour at room temperature, then incubated with the AS160 (pS642) antibody for two hours at room temperature in 3% BSA-TBST buffer, following prior incubation with: no peptide (1, 5-7), the non-phosphopeptide corresponding to the phosphopeptide immunogen (2), a generic phosphothreonine-containing peptide (3), or the phosphopeptide immunogen (4). After washing, the membrane was incubated with goat F (ab')2 anti-rabbit IgG HRP conjugate (Product # ALI4404) and signals were detected using the Pierce SuperSignal™ method. The data show that only the phosphopeptide corresponding to AS160 (pS642) blocks the signal. The data also show that the signal was blocked in cells overexpressing site-directed mutant T642A, indicating the antibody is phosphorylation site-specific. Note: The COS cells overexpressing AS160 were a generous gift from Dr. Gustav Lienhard (Dartmouth Medical School).

Product Details

Tested Applications

Dilution

Western Blot (WB)

Assay Dependent

Published Applications

Western Blot (WB)

See 8 publications below

Immunofluorescence (IF)

See 1 publication below

Immunohistochemistry (IHC)

See 1 publication below

Product Specifications

Species Reactivity

Human, Mouse

Published species

Human, Mouse

Host / Isotype

Rabbit / IgG

Class

Polyclonal

Type

Antibody

Immunogen

The antiserum was produced against a chemically synthesized phosphopeptide derived from the region of human AS160 that contains threonine 642. The sequence is conserved in mouse.

Conjugate

Unconjugated

Form

Liquid

Purification

Antigen affinity chromatography

Storage buffer

Dulbecco's PBS, pH 7.3, with 1mg/mL BSA, 50% glycerol

Contains

0.05% sodium azide

Storage conditions

-20°C

RRID

AB_2533564

Target Information

TBC1D4, also known as the Akt substrate AS160, was initially identified as an Akt substrate containing a GTPase-activating domain that regulates GLUT4 trafficking, with activation following insulin stimulation. TBC1D4 truncations in humans is a major cause of dominant inherited insulin resistance. The loss of TBC1D4 results in the accumulation of GLUT4 in compartments that are primed for fusion in basal adipocytes.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Bioinformatics

Protein Aliases: Acrg embryonic lethality minimal region ortholog; Akt substrate of 160 kDa; AS160; BUB2; CDC16) domain-containing protein; TBC (Tre-2; TBC (Tre-2, BUB2, CDC16) domain-containing protein; TBC1 domain family member 4; TBC1 domain family, member 4

Gene Aliases: 5930406J04Rik; A930035N22; AS160; AV295684; KIAA0603; NIDDM5; TBC1D4

UniProt ID: (Human) O60343, (Mouse) Q3TAQ5

Entrez Gene ID: (Human) 9882, (Mouse) 210789

Molecular Function: G-protein modulator enzyme modulator hydrolase