Invitrogen

Phospho-AS160 (Thr642) Polyclonal Antibody

Advanced Verification

This Antibody was verified by Cell treatment to ensure that the antibody binds to the antigen stated.

8 Published Figures

12 References

View all (28) 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. 7. Effects of 6-MP treatment on protein levels of phosphorylated and total Akt substrate of 160 kDa (AS160) and on mRNA level of AS160 in L6 skeletal muscle cells. A : dose effects of 6-MP on protein levels of phosphorylated and total AS160 compared with NR4A3 protein expression in L6 myotubes. L6 myotubes were treated with DMSO vehicle control (0 muM) or 5-75 muM 6-MP for 24 h and further incubated in basal or 100 nM insulin for 60 min at 37degC. Total proteins were then separated by electrophoresis and analyzed by immunoblotting with antibodies against phosphorylated AS160 (p-AS160 Thr 642 ), total AS160, and NR4A3. beta-Actin was measured as an internal control. Representative blots are shown. The relative protein signals were quantified densitometrically, and data shown are means +- SD ( n >= 3). * P < 0.05 and ** P < 0.01 compared with basal control (0 muM); # P < 0.05 and ## P < 0.01 compared with insulin-stimulated control (0 muM). B : dose effects of 6-MP on mRNA level of AS160 in L6 myotubes by RT-PCR. L6 myotubes were treated with DMSO control (0 muM) or 5-75 muM 6-MP for 24 h. Total RNAs were extracted and then RT-PCR and gel electrophoresis were conducted for the analysis of mRNA level of AS160. 18S was measured as an internal control. Representative gel images are shown. Relative mRNAs were quantified densitometrically, and data shown are means +- SD ( n >= 2). {TM}

Product Details

44-1071G

Applications	Tested Dilution	Publications
Western Blot (WB)	1:1,000	View 11 publications 11 publications
Immunohistochemistry (IHC)	-	View 1 publication 1 publication

Product Specifications
Species Reactivity	Human, Mouse
Published species	Human, Mouse, Rat
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. View immunogen
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.

Product References

Inhibition of AMPK activity in response to insulin in adipocytes: involvement of AMPK pS485, PDEs, and cellular energy levels.

American journal of physiology. Endocrinology and metabolism

Kopietz F,Rupar K,Berggreen C,Säll J,Vertommen D,Degerman E,Rider MH,Göransson O

44-1071G was used in Western Blot to improve the understanding of how insulin regulates AMP-activated protein kinase activity, and with that, de novo fatty acid synthesis, in adipose tissue.

Tue Sep 01 00:00:00 EDT 2020

Glucose Sensing by Skeletal Myocytes Couples Nutrient Signaling to Systemic Homeostasis.

Molecular cell

Meng ZX,Gong J,Chen Z,Sun J,Xiao Y,Wang L,Li Y,Liu J,Xu XZS,Lin JD

44-1071G was used in Western Blotting to uncover an unexpected mechanism of glucose sensing in skeletal myocytes that contributes to homeostasis and therapeutic action.

Thu May 04 00:00:00 EDT 2017

Skeletal Muscle TRIB3 Mediates Glucose Toxicity in Diabetes and High- Fat Diet-Induced Insulin Resistance.

Diabetes

Zhang W,Wu M,Kim T,Jariwala RH,Garvey WJ,Luo N,Kang M,Ma E,Tian L,Steverson D,Yang Q,Fu Y,Garvey WT

44-1071G was used in Western Blotting to demonstrate that TRIB3 has a pathophysiological role in diabetes and a physiological role in metabolism.

Mon Aug 01 00:00:00 EDT 2016

PKB-Mediated Thr649 Phosphorylation of AS160/TBC1D4 Regulates the R-Wave Amplitude in the Heart.

PloS one

Quan C,Xie B,Wang HY,Chen S

44-1071G was used in western blot to determine that PKB-mediated phosphorylation of AS160/TBC1D4 is one of the major PAS-binding signals in the heart in response to insulin.

Mon Feb 08 00:00:00 EST 2016

TBC1D1 reduces palmitate oxidation by inhibiting β-HAD activity in skeletal muscle.

American journal of physiology. Regulatory, integrative and comparative physiology

Maher AC,McFarlan J,Lally J,Snook LA,Bonen A

44-1071G was used in Western Blotting to demonstrate that TBC1D1-mediated reduction of muscle fatty acid oxidation appears to occur via inhibition of Β-HAD activity.

Sat Nov 01 00:00:00 EDT 2014

6-Mercaptopurine augments glucose transport activity in skeletal muscle cells in part via a mechanism dependent upon orphan nuclear receptor NR4A3.

American journal of physiology. Endocrinology and metabolism

Liu Q,Zhu X,Xu L,Fu Y,Garvey WT

Published figure using Phospho-AS160 (Thr642) polyclonal antibody (Product # 44-1071G) in Western Blot

Fri Nov 01 00:00:00 EDT 2013

Thr649Ala-AS160 knock-in mutation does not impair contraction/AICAR-induced glucose transport in mouse muscle.

American journal of physiology. Endocrinology and metabolism

Ducommun S,Wang HY,Sakamoto K,MacKintosh C,Chen S

44-1071G was used in western blot to determine the role of AS60-Thr(649) phosphorylation/4-3-3 binding in mediating glucose uptake.

Tue May 15 00:00:00 EDT 2012

Recovered insulin response by 2 weeks of leptin administration in high-fat fed rats is associated with restored AS160 activation and decreased reactive lipid accumulation.

American journal of physiology. Regulatory, integrative and comparative physiology

Stefanyk LE,Gulli RA,Ritchie IR,Chabowski A,Snook LA,Bonen A,Dyck DJ

Fri Jul 01 00:00:00 EDT 2011

Loss of 50% of excess weight using a very low energy diet improves insulin-stimulated glucose disposal and skeletal muscle insulin signalling in obese insulin-treated type 2 diabetic patients.

Diabetologia

Jazet IM,Schaart G,Gastaldelli A,Ferrannini E,Hesselink MK,Schrauwen P,Romijn JA,Maassen JA,Pijl H,Ouwens DM,Meinders AE

Fri Feb 01 00:00:00 EST 2008

Insulin-mediated phosphorylation of the proline-rich Akt substrate PRAS40 is impaired in insulin target tissues of high-fat diet-fed rats.

Diabetes

Nascimento EB,Fodor M,van der Zon GC,Jazet IM,Meinders AE,Voshol PJ,Vlasblom R,Baan B,Eckel J,Maassen JA,Diamant M,Ouwens DM

Fri Dec 01 00:00:00 EST 2006

Insulin-mediated phosphorylation of the proline-rich Akt substrate PRAS40 is impaired in insulin target tissues of high-fat diet-fed rats.

Diabetes

Nascimento EB,Fodor M,van der Zon GC,Jazet IM,Meinders AE,Voshol PJ,Vlasblom R,Baan B,Eckel J,Maassen JA,Diamant M,Ouwens DM

Fri Dec 01 00:00:00 EST 2006

Insulin-mediated phosphorylation of the proline-rich Akt substrate PRAS40 is impaired in insulin target tissues of high-fat diet-fed rats.

Diabetes

Nascimento EB,Fodor M,van der Zon GC,Jazet IM,Meinders AE,Voshol PJ,Vlasblom R,Baan B,Eckel J,Maassen JA,Diamant M,Ouwens DM

Fri Dec 01 00:00:00 EST 2006

Distinct signals regulate AS160 phosphorylation in response to insulin, AICAR, and contraction in mouse skeletal muscle.

Diabetes

Kramer HF,Witczak CA,Fujii N,Jessen N,Taylor EB,Arnolds DE,Sakamoto K,Hirshman MF,Goodyear LJ

44-1071G was used in western blot to study the relationships among Akt, AMPK alpha2, AS60 phosphorylation, insulin, and AICAR.

Sat Jul 01 00:00:00 EDT 2006

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: scaffold/adaptor protein GTPase-activating protein