Overview

  • Product nameAnti-TBC1D4 (phospho T642) antibody
    See all TBC1D4 primary antibodies
  • Description
    Sheep polyclonal to TBC1D4 (phospho T642)
  • Tested applicationsSuitable for: WB, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide: CRRRAHTPFSHPP conjugated to KLH, corresponding to amino acids 637-647 of Human TBC1D4

  • Positive control
    • 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

Properties

  • FormLiquid
  • 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 bufferPreservative: 0.05% Sodium Azide
    Constituents: 150mM Sodium chloride, 10mM HEPES, pH 7.5
  • Concentration information loading...
  • PurityImmunogen affinity purified
  • ClonalityPolyclonal
  • IsotypeIgG
  • Research areas

Applications

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.

Application Abreviews Notes
WB Use a concentration of 2 µg/ml. Predicted molecular weight: 147 kDa.
ICC/IF Use a concentration of 10 µg/ml.

Target

  • 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.
  • Post-translational
    modifications
    Phosphorylated 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
  • Database links
  • Alternative names
    • Acrg embryonic lethality (mouse) minimal region ortholog antibody
    • Acrg embryonic lethality minimal region ortholog antibody
    • Acrg embryonic lethality mouse minimal region ortholog antibody
    • Akt substrate of 160 kDa antibody
    • AS 160 antibody
    • AS160 antibody
    • BUB2 antibody
    • CDC16 antibody
    • KIAA0603 antibody
    • TBC (Tre 2 BUB2 CDC16) domain containing protein antibody
    • TBC Tre 2 BUB2 CDC16 domain containing protein antibody
    • TBC1 D4 antibody
    • TBC1 domain family member 4 antibody
    • Tbc1d4 antibody
    • TBCD4_HUMAN antibody
    • Tre-2 antibody
    see all

Anti-TBC1D4 (phospho T642) antibody images

  • All lanes : Anti-TBC1D4 (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
  • 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.

References for Anti-TBC1D4 (phospho T642) antibody (ab65753)

This product has been referenced in:
  • 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) »
  • Schweitzer GG  et al. Sustained postexercise increases in AS160 Thr642 and Ser588 phosphorylation in skeletal muscle without sustained increases in kinase phosphorylation. J Appl Physiol 113:1852-61 (2012). WB ; Rat . Read more (PubMed: 22936728) »

See all 2 Publications for this product

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Rat Tissue lysate - other (Skeletal Muscle)
Loading amount 60 µg
Specification Skeletal Muscle
Treatment incubated with or without 100uUnits of insulin
Gel Running Conditions Reduced Denaturing (8%)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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Submitted Dec 20 2011

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"