Product nameAnti-AS160 (phospho T642) antibody
See all AS160 primary antibodies
DescriptionSheep polyclonal to AS160 (phospho T642)
Tested applicationsSuitable for: WB, ICC/IFmore details
Species reactivityReacts with: Mouse, Rat, Human
- Tagged AS160 wild type, S588A mutant, and T642A mutant over-expressed transiently in HEK293 cells, basal or with 1 µM insulin treatment for 15 minutes. IF: HeLa cell line
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Store at -20°C or -80°C. Avoid freeze / thaw cycle.
Storage bufferpH: 7.50
Preservative: 0.05% Sodium azide
Constituents: 0.238% HEPES, 0.87% Sodium chloride
Concentration information loading...
PurityImmunogen affinity purified
Our Abpromise guarantee covers the use of ab65753 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||Use a concentration of 2 µg/ml. Predicted molecular weight: 147 kDa.|
|ICC/IF||Use a concentration of 10 µg/ml.|
FunctionMay act as a GTPase-activating protein for RAB2A, RAB8A, RAB10 and RAB14. Isoform 2 promotes insulin-induced glucose transporter SLC2A4/GLUT4 translocation at the plasma membrane, thus increasing glucose uptake.
Tissue specificityWidely expressed. Isoform 2 is the highest overexpressed in most tissues. Isoform 1 is highly expressed in skeletal muscle and heart, but was not detectable in the liver nor in adipose tissue. Isoform 2 is strongly expressed in adrenal and thyroid gland, and also in lung, kidney, colon, brain and adipose tissue. Isoform 2 is moderately expressed in skeletal muscle. Expressed in pancreatic Langerhans islets, including beta cells (at protein level). Expression is decreased by twofold in pancreatic islets in type 2 diabetes patients compared to control subjects. Up-regulated in T cells from patients with atopic dermatitis.
Sequence similaritiesContains 2 PID domains.
Contains 1 Rab-GAP TBC domain.
modificationsPhosphorylated by AKT1; insulin-induced.
Insulin-stimulated phosphorylation is required for SLC2A4/GLUT4 translocation.
Physiological hyperinsulinemia increases phosphorylation in skeletal muscle. Insulin-stimulated phosphorylation is reduced by 39% in type 2 diabetic patients.
Cellular localizationCytoplasm. Isoform 2 shows a cytoplasmic perinuclear localization in a myoblastic cell line in resting and insulin-stimulated cells.
- Information by UniProt
- Acrg embryonic lethality (mouse) minimal region ortholog antibody
- Acrg embryonic lethality minimal region ortholog antibody
- Acrg embryonic lethality mouse minimal region ortholog antibody
All lanes : Anti-AS160 (phospho T642) antibody (ab65753) at 2 µg/ml
Lane 1 : AS160 wild type (WT)
Lane 2 : AS160 wild type (WT) with (+) 1 µM insulin treatment for 15 minutes
Lane 3 : S588A mutant (S588A)
Lane 4 : S588A mutant (S588A) with (+) 1 µM insulin treatment for 15 minutes
Lane 5 : T642A mutant (T642A)
Lane 6 : T642A mutant (T642A) with (+) 1 µM insulin treatment for 15 minutes
Predicted band size: 147 kDa
Western blot analysis of immunoprecipitated Flagtagged AS160 wild type (WT), S588A mutant (S588A), and T642A mutant (T642A) over-expressed transiently in HEK293 cells, basal (-) or with (+) 1 µM insulin treatment for 15 minutes.
ICC/IF image of ab65753 stained HeLa cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal donkey serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab65753, 10µg/ml) overnight at +4°C. The secondary antibody (green) was ab96939, DyLight® 488 donkey anti-sheep IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
This product has been referenced in:
- Yang GS et al. mTOR is involved in stroke-induced seizures and the anti-seizure effect of mild hypothermia. Mol Med Rep 17:5821-5829 (2018). Read more (PubMed: 29484389) »
- Inazuka F et al. Muscle-specific Knock-out of NUAK Family SNF1-like Kinase 1 (NUAK1) Prevents High Fat Diet-induced Glucose Intolerance. J Biol Chem 287:16379-89 (2012). WB ; Mouse . Read more (PubMed: 22418434) »