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    as160-phospho-t642-antibody-epr27332-ab131214.pdf

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RecombinantRabMAb

Recombinant Anti-AS160 (phospho T642) antibody [EPR2733(2)] (ab131214)

  • Datasheet
  • SDS
Reviews (2) Submit a question References (1)

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Western blot - Anti-AS160 (phospho T642) antibody [EPR2733(2)] (ab131214)
  • Dot Blot - Anti-AS160 (phospho T642) antibody [EPR2733(2)] (ab131214)
  • Western blot - Anti-AS160 (phospho T642) antibody [EPR2733(2)] (ab131214)
  • OI-RD Scanning - Anti-AS160 (phospho T642) antibody [EPR2733(2)] (ab131214)
  • Anti-AS160 (phospho T642) antibody [EPR2733(2)] (ab131214)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR2733(2)] to AS160 (phospho T642)
  • Suitable for: Dot blot, WB
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Carrier Free

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Overview

  • Product name

    Anti-AS160 (phospho T642) antibody [EPR2733(2)]
    See all AS160 primary antibodies
  • Description

    Rabbit monoclonal [EPR2733(2)] to AS160 (phospho T642)
  • Host species

    Rabbit
  • Tested applications

    Suitable for: Dot blot, WBmore details
    Unsuitable for: Flow Cyt,ICC/IF,IHC-P or IP
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Synthetic peptide within Human AS160 (phospho T642). The exact sequence is proprietary.
    Database link: O60343

  • Positive control

    • WB: HEK-293 grown in serum free media overnight, then treated with 100nM Calyculin A (ab141784) for 50min and then 100ng/ml Insulin was added for the last 20min, whole cell lysate and 293T cell lysate - insulin-treated. Dot Blot: AS160 (phospho T642) phospho peptide.
  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C.
  • Dissociation constant (KD)

    KD = 6.40 x 10 -11 M
    Learn more about KD
  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 9% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA, 50% Tissue culture supernatant
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR2733(2)
  • Isotype

    IgG
  • Research areas

    • Signal Transduction
    • Signaling Pathway
    • G Protein Signaling
    • Small G Proteins
    • Other

Associated products

  • Alternative Versions

    • Anti-AS160 (phospho T642) antibody [EPR2733(2)] - BSA and Azide free (ab218844)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Related Products

    • Calyculin A, protein phosphatase inhibitor (ab141784)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab131214 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
Dot blot
1/1000.
WB (2)
1/1000 - 1/10000. Predicted molecular weight: 146 kDa.
Notes
Dot blot
1/1000.
WB
1/1000 - 1/10000. Predicted molecular weight: 146 kDa.
Application notes
Is unsuitable for Flow Cyt,ICC/IF,IHC-P or IP.

Target

  • Function

    May 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 specificity

    Widely 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 similarities

    Contains 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 localization

    Cytoplasm. Isoform 2 shows a cytoplasmic perinuclear localization in a myoblastic cell line in resting and insulin-stimulated cells.
  • Target information above from: UniProt accession O60343 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 9882 Human
    • Omim: 612465 Human
    • SwissProt: O60343 Human
    • Unigene: 210891 Human
    • 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
      • NIDDM5 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

    Images

    • Western blot - Anti-AS160 (phospho T642) antibody [EPR2733(2)] (ab131214)
      Western blot - Anti-AS160 (phospho T642) antibody [EPR2733(2)] (ab131214)
      All lanes : Anti-AS160 (phospho T642) antibody [EPR2733(2)] (ab131214) at 1.12 µg/ml

      Lane 1 : HEK-293 (human embryonic kidney epithelial cell) grown in serum free media overnight whole cell lysate
      Lane 2 : HEK-293 grown in serum free media overnight, then treated with 100nM Calyculin A (ab141784) for 50min and then 100ng/ml Insulin was added for the last 20min, whole cell lysate
      Lane 3 : HEK-293 grown in serum free media overnight, then treated with 100nM Calyculin A (ab141784) for 50min and then 100ng/ml Insulin was added for the last 20min, whole cell lysate. Then the membrane was incubated with alkaline phosphatase

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size: 146 kDa



      Blocking and diluting buffer: 5% NFDM/TBST. 

    • Dot Blot - Anti-AS160 (phospho T642) antibody [EPR2733(2)] (ab131214)
      Dot Blot - Anti-AS160 (phospho T642) antibody [EPR2733(2)] (ab131214)

      Dot blot analysis of AS160 (phospho T642) phospho peptide (Lane 1) and AS160 non-phospho peptide (Lane 2) labelling AS160 (phospho T642) phospho peptide with ab131214 at a dilution of 1:1000 dilution (1.12μg/ml). A Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) was used as the secondary antibody at a dilution of 1:20,000 dilution.

      Blocking and dilution buffer: 5% NFDM/TBST.

    • Western blot - Anti-AS160 (phospho T642) antibody [EPR2733(2)] (ab131214)
      Western blot - Anti-AS160 (phospho T642) antibody [EPR2733(2)] (ab131214)
      All lanes : Anti-AS160 (phospho T642) antibody [EPR2733(2)] (ab131214) at 1/1000 dilution

      Lane 1 : 293T cell lysate - untreated
      Lane 2 : 293T cell lysate - insulin-treated

      Predicted band size: 146 kDa



      Bottom pannel shows detection of total AS160 with a general anti AS160 antibody.

    • OI-RD Scanning - Anti-AS160 (phospho T642) antibody [EPR2733(2)] (ab131214)
      OI-RD Scanning - Anti-AS160 (phospho T642) antibody [EPR2733(2)] (ab131214)
      Equilibrium disassociation constant (KD)
      Learn more about KD

      Click here to learn more about KD
    • Anti-AS160 (phospho T642) antibody [EPR2733(2)] (ab131214)
      Anti-AS160 (phospho T642) antibody [EPR2733(2)] (ab131214)

    Protocols

    • Western blot protocols

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (1)

    Publishing research using ab131214? Please let us know so that we can cite the reference in this datasheet.

    ab131214 has been referenced in 1 publication.

    • Tobias IS  et al. Sex and fiber type independently influence AMPK, TBC1D1, and TBC1D4 at rest and during recovery from high-intensity exercise in humans. J Appl Physiol (1985) 128:350-361 (2020). PubMed: 31895596

    Customer reviews and Q&As

    Show All Reviews Q&A
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    1-2 of 2 Abreviews or Q&A

    Western blot abreview for Anti-AS160 (phospho T642) antibody [EPR2733(2)]

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Mouse Tissue lysate - whole (brown adipose tissue)
    Gel Running Conditions
    Reduced Denaturing (10 %)
    Loading amount
    60 µg
    Treatment
    fasted o/n and refed 2hrs
    Specification
    brown adipose tissue
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 23°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Aug 23 2021

    Western blot abreview for Anti-AS160 (phospho T642) antibody [EPR2733(2)]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Tissue lysate - whole (Muscle fibers)
    Gel Running Conditions
    Reduced Denaturing (66-440 kDa separation matrix)
    Loading amount
    5 µg
    Specification
    Muscle fibers
    Blocking step
    ProteinSimple Blocking Buffer as blocking agent for 5 minute(s) · Concentration: 100% · Temperature: 24°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Irene Tobias

    Verified customer

    Submitted Mar 19 2018

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