Invitrogen

Phospho-AS160 (Thr642) Polyclonal Antibody

8 Published Figures

12 References

View all (27) AS160 antibodies

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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).

Phospho-AS160 (Thr642) Antibody — Fig 4 Expression and phosphorylation of AS160 and TBC1D1 in the infarct heart. Immunoblotting analyses of phosphorylation of AS160-Thr 649 and PKB-Ser 473 , and total AS160, TBC1D1, PKB, GLUT4 and cTnT in the lysates (40 mug) of the infarct zone (I), border zone (B) and remote zone (R) of mouse heart 4 weeks after myocardial infarction. Flotillin-1 was used as a loading control. A, representative blots. B, quantitation of TBC1D1 signals, n = 4. Asterisk indicates p < 0.05 (infarct zone versus remote zone).

Product Details

Applications

Tested Dilution

Publications

Western Blot (WB)

1:1000

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Immunofluorescence (IF)

View 1 publication 1 publication

Immunohistochemistry (IHC)

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Product Specifications

Species

Human, Mouse

Published species

Human, Mouse

Expression System

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

AS160 (TBC1D4) is a GTPase-activiting protein for RAB2A, RAB8A, RAB10, and RAB14. The isoform 2 version of this protein promotes the insulin-induced glucose transporter SLC2A4/GLUT4 translocation at the plasma membrane. This movement increases glucose uptake. Mutations associated with the gene cause Diabetes mellitus, non-insulin-dependent, 5 (NIDDM5).

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

Bioinformatics

Protein Aliases: Akt substrate of 160 kDa; AS160; DKFZp779C0666; OTTHUMP00000040818; RP11-159J2.1; RP11-159J2.3; TBC (Tre-2, BUB2, CDC16) domain-containing protein; TBC Tre-2; TBC1 domain family member 4; TBC1 domain family, member 4

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

UniProt ID: (Human) O60343, (Mouse) Q8BYJ6

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

Molecular Function: G-protein modulator enzyme modulator hydrolase