Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-LRP6 antibody [EPR22910-39] (ab231779)

Overview

  • Product name

    Anti-LRP6 antibody [EPR22910-39]
    See all LRP6 primary antibodies
  • Description

    Rabbit monoclonal [EPR22910-39] to LRP6
  • Host species

    Rabbit
  • Tested applications

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

    Reacts with: Human
  • Immunogen

    Synthetic peptide within Human LRP6 aa 1500-1600. The exact sequence is proprietary.
    Database link: O75581

  • Positive control

    • WB: HAP1 (WT), HeLa, HepG2 and HEK-293T whole cell lysates.
  • General notes

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

    This product is a recombinant rabbit monoclonal antibody.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer

    Preservative: 0.01% Sodium azide
    Constituents: PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR22910-39
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab231779 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 1/1000. Detects a band of approximately 180-210 kDa (predicted molecular weight: 180 kDa).
  • Application notes
    Is unsuitable for Flow Cyt,ICC/IF,IHC-P or IP.
  • Target

    • Function

      Component of the Wnt-Fzd-LRP5-LRP6 complex that triggers beta-catenin signaling through inducing aggregation of receptor-ligand complexes into ribosome-sized signalsomes. Cell-surface coreceptor of Wnt/beta-catenin signaling, which plays a pivotal role in bone formation. The Wnt-induced Fzd/LRP6 coreceptor complex recruits DVL1 polymers to the plasma membrane which, in turn, recruits the AXIN1/GSK3B-complex to the cell surface promoting the formation of signalsomes and inhibiting AXIN1/GSK3-mediated phosphorylation and destruction of beta-catenin. Required for posterior patterning of the epiblast during gastrulation.
    • Tissue specificity

      Widely co-expressed with LRP5 during embryogenesis and in adult tissues.
    • Involvement in disease

      Defects in LRP6 are the cause of autosomal dominant coronary artery disease type 2 (ADCAD2) [MIM:610947].
    • Sequence similarities

      Belongs to the LDLR family.
      Contains 4 EGF-like domains.
      Contains 3 LDL-receptor class A domains.
      Contains 20 LDL-receptor class B repeats.
    • Domain

      The YWTD-EGF-like domains 1 and 2 are required for the interaction with Wnt-frizzled complex. The YWTD-EGF-like domains 3 and 4 are required for the interaction with DKK1.
      The PPPSP motifs play a central role in signal transduction by being phosphorylated, leading to activate the Wnt signaling pathway.
    • Post-translational
      modifications

      Dual phosphorylation of cytoplasmic PPPSP motifs sequentially by GSK3 and CK1 is required for AXIN1-binding, and subsequent stabilization and activation of beta-catenin via preventing GSK3-mediated phosphorylation of beta-catenin. Phosphorylated, in vitro, by GRK5/6 within and outside the PPPSP motifs. Phosphorylation at Ser-1490 by CDK14 during G2/M phase leads to regulation of the Wnt signaling pathway during the cell cycle. Phosphorylation by GSK3B is induced by RPSO1 binding and inhibited by DKK1. Phosphorylated, in vitro, by casein kinase I on Thr-1479.
      Undergoes gamma-secretase-dependent regulated intramembrane proteolysis (RIP). The extracellular domain is first released by shedding, and then, through the action of gamma-secretase, the intracellular domain (ICD) is released into the cytoplasm where it is free to bind to GSK3B and to activate canonical Wnt signaling.
      Palmitoylation on the two sites near the transmembrane domain leads to release of LRP6 from the endoplasmic reticulum.
      Mono-ubiquitinated which retains LRP6 in the endoplasmic reticulum.
      N-glycosylation is required for cell surface location.
    • Cellular localization

      Membrane. Endoplasmic reticulum. On Wnt signaling, undergoes a cycle of caveolin- or clathrin-mediated endocytosis and plasma membrane location. Released from the endoplasmic reticulum on palmitoylation. Mono-ubiquitination retains it in the endoplasmic reticulum in the absence of palmitoylation. On Wnt signaling, phosphorylated, aggregates and colocalizes with AXIN1 and GSK3B at the plasma membrane in LRP6-signalsomes. Chaperoned to the plasma membrane by MESD.
    • Information by UniProt
    • Database links

    • Alternative names

      • ADCAD2 antibody
      • C030016K15Rik antibody
      • Cd antibody
      • FLJ90062 antibody
      • FLJ90421 antibody
      • LDL receptor related protein 6 antibody
      • Low density lipoprotein receptor related protein 6 antibody
      • Low-density lipoprotein receptor-related protein 6 antibody
      • LRP-6 antibody
      • LRP6 antibody
      • LRP6_HUMAN antibody
      • OTTHUMP00000238979 antibody
      • OTTHUMP00000238980 antibody
      • OTTHUMP00000238982 antibody
      • STHAG7 antibody
      see all

    Images

    • All lanes : Anti-LRP6 antibody [EPR22910-39] (ab231779) at 1/1000 dilution

      Lane 1 : Wild-type HAP1 whole cell lysate
      Lane 2 : LRP6 knockout HAP1 whole cell lysate
      Lane 3 : HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate
      Lane 4 : HepG2 (human hepatocellular carcinoma epithelial cell), whole cell lysate
      Lane 5 : 293T (human embryonic kidney epithelial cell), whole cell lysate

      Lysates/proteins at 20 µg per lane.

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

      Predicted band size: 180 kDa
      Observed band size: 180-210 kDa
      why is the actual band size different from the predicted?



      The molecular weight observed is consistent with what has been described in the literature (PMID: 16989816, 16263759, 28341812).  ab231779 was shown to specifically react with LRP6 in wild-type HAP1 cells as signal was lost in LRP6 knockout cells. Wild-type and LRP6 knockout samples were subjected to SDS-PAGE. ab231779 and ab129002 (Rabbit anti-Vinculin loading control) were incubated 1 hour at room temperature at 1/1000 dilution and 1/200,000 dilution respectively. Blots were developed with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) secondary antibody at 1/20,000 dilution for 1 hour at room temperature before imaging. The blot was developed on a BIO-RAD® ChemiDoc™ instrument using the ECL technique.  This blot was developed using a higher sensitivity ECL substrate.

      Blocking/Dilution buffer: 5% NFDM/TBST.

      Exposure times: Lanes 1-2: 3 mins; Lanes 3: 26 secs; Lanes 4-5: 3 mins.

    • All lanes : Anti-LRP6 antibody [EPR22910-39] (ab231779) at 1/1000 dilution

      Lane 1 : His-tagged human LRP5 recombinant protein, 10 ng
      Lane 2 : His-tagged human LRP6 recombinant protein, 10 ng

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

      Predicted band size: 180 kDa
      Observed band size: 18,36 kDa why is the actual band size different from the predicted?


      Exposure time: 1 second


      The 36 KD band observed is consistent with the dimer of human LRP6 recombinant protein (aa 1476-1600).

    References

    ab231779 has not yet been referenced specifically in any publications.

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