Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-ApoER2 antibody [EPR3326] (ab108208)

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Western blot - Anti-ApoER2 antibody [EPR3326] (ab108208)
  • Immunoprecipitation - Anti-ApoER2 antibody [EPR3326] (ab108208)
  • Western blot - Anti-ApoER2 antibody [EPR3326] (ab108208)
  • Western blot - Anti-ApoER2 antibody [EPR3326] (ab108208)
  • Anti-ApoER2 antibody [EPR3326] (ab108208)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR3326] to ApoER2
  • Suitable for: WB, IP
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Carrier Free

Overview

  • Product name

    Anti-ApoER2 antibody [EPR3326]
    See all ApoER2 primary antibodies

  • Description

    Rabbit monoclonal [EPR3326] to ApoER2

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, IPmore details
    Unsuitable for: ICC or IHC-P

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • SH-SY5Y, C6, Neuro-2a, Human fetal brain, Mouse brain, and Rat brain lysates,

  • 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.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

Properties

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab108208 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)
1/1000 - 1/10000. Predicted molecular weight: 106 kDa.
IP
1/10 - 1/100.
Notes
WB
1/1000 - 1/10000. Predicted molecular weight: 106 kDa.
IP
1/10 - 1/100.
  • Application notes
    Is unsuitable for ICC or IHC-P.

    • Target

    • Function

      Cell surface receptor for Reelin (RELN) and apolipoprotein E (apoE)-containing ligands. LRP8 participates in transmitting the extracellular Reelin signal to intracellular signaling processes, by binding to DAB1 on its cytoplasmic tail. Reelin acts via both the VLDL receptor (VLDLR) and LRP8 to regulate DAB1 tyrosine phosphorylation and microtubule function in neurons. LRP8 has higher affinity for Reelin than VLDLR. LRP8 is thus a key component of the Reelin pathway which governs neuronal layering of the forebrain during embryonic brain development. Binds the endoplasmic reticulum resident receptor-associated protein (RAP). Binds dimers of beta 2-glycoprotein I and may be involved in the suppression of platelet aggregation in the vasculature. Highly expressed in the initial segment of the epididymis, where it affects the functional expression of clusterin and phospholipid hydroperoxide glutathione peroxidase (PHGPx), two proteins required for sperm maturation. May also function as an endocytic receptor.

    • Tissue specificity

      Expressed mainly in brain and placenta. Also expressed in platelets and megakaryocytic cells. Not expressed in the liver.

    • Involvement in disease

      Defects in LRP8 are a cause of myocardial infarction type 1 (MCI1) [MIM:608446]. A condition defined by the irreversible necrosis of heart muscle secondary to prolonged ischemia.

    • Sequence similarities

      Belongs to the LDLR family.
      Contains 2 EGF-like domains.
      Contains 7 LDL-receptor class A domains.
      Contains 5 LDL-receptor class B repeats.

    • Domain

      The cytoplasmic domain is involved in the binding of DAB1 and in the recruitment of JNK-interacting proteins. Isoforms, which lack part of the cytoplasmic domain, are unable to recruit members of the family of JNK interacting proteins (JIP) to the cytoplasmic tail.

    • Post-translational
      modifications

      O-glycosylated. Some alternatively spliced isoforms lack the O-linked sugar domain.
      Undergoes sequential, furin and gamma-secretase dependent, proteolytic processing, resulting in the extracellular release of the entire ligand-binding domain as a soluble polypeptide and in the intracellular domain (ICD) release into the cytoplasm. The gamma-secretase-dependent proteolytical processing occurs after the bulk of the extracellular domain has been shed, in a furin-dependent manner, in alternatively spliced isoforms carrying the furin cleavage site. Hypoglycosylation (mainly hypo-O-glycosylation) leads to increased extracellular cleavage, which in turn results in accelerating release of the intracellular domain (ICD) by the gamma-secretase. The resulting receptor fragment is able to inhibit Reelin signaling and in particular the Reelin-induced DAB1 phosphorylation.
      Tyrosine phosphorylated upon apoE binding.
      Ubiquitinated by MYLIP leading to degradation.

    • Cellular localization

      Cell membrane. Secreted. Isoforms that contain the exon coding for a furin-type cleavage site are proteolytically processed, leading to a secreted receptor fragment.
    • Information by UniProt

    • Database links

      • Alternative names

        • APOER2 antibody
        • Apolipoprotein E receptor 2 antibody
        • low density lipoprotein receptor-related protein 8 antibody
        • Low-density lipoprotein receptor-related protein 8 antibody
        • LRP-8 antibody
        • LRP8 antibody
        • LRP8_HUMAN antibody
        see all

      Images

      • Western blot - Anti-ApoER2 antibody [EPR3326] (ab108208)
        Western blot - Anti-ApoER2 antibody [EPR3326] (ab108208)
        All lanes : Anti-ApoER2 antibody [EPR3326] (ab108208) at 1/1000 dilution

        Lane 1 : Wild-type HAP1 whole cell lysate
        Lane 2 : ApoER2 knockout HAP1 whole cell lysate

        Lysates/proteins at 40 µg per lane.

        Predicted band size: 106 kDa



        Lanes 1 - 2: Merged signal (red and green). Green - ab108208 observed at 106 kDa. Red - loading control, ab9484, observed at 37 kDa.

        ab108208 was shown to specifically react with ApoER2 in wild-type HAP1 cells as signal was lost in LRP8 knockout cells. Wild-type and ApoER2 knockout samples were subjected to SDS-PAGE. Ab108208 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

      • Immunoprecipitation - Anti-ApoER2 antibody [EPR3326] (ab108208)
        Immunoprecipitation - Anti-ApoER2 antibody [EPR3326] (ab108208)

        ab108208 (purified) at 1/40 immunoprecipitating ApoER2 in SH-SY5Y (human neuroblastoma) whole cell lysate.

        Lane 1 (input): SH-SY5Y whole cell lysate 10ug

        Lane 2 (+): ab108208 + SH-SY5Y whole cell lysate

        Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab108208 in SH-SY5Y whole cell lysate

        For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution.

        Blocking buffer and concentration: 5% NFDM/TBST.

        Diluting buffer and concentration: 5% NFDM /TBST.

      • Western blot - Anti-ApoER2 antibody [EPR3326] (ab108208)
        Western blot - Anti-ApoER2 antibody [EPR3326] (ab108208)
        All lanes : Anti-ApoER2 antibody [EPR3326] (ab108208) at 1/10000 dilution (purified)

        Lane 1 : SH-SY5Y (human neuroblastoma) whole cell lysate
        Lane 2 : Human fetal brain tissue lysate
        Lane 3 : Mouse brain tissue lysate
        Lane 4 : C6 (rat glioma) whole cell lysate
        Lane 5 : Neuro-2a (mouse neuroblastoma) whole cell lysate

        Lysates/proteins at 20 µg per lane.

        Secondary
        All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/2000 dilution

        Predicted band size: 106 kDa
        Observed band size: 130-170 kDa why is the actual band size different from the predicted?



        Blocking and diluting buffer 5% NFDM/TBST.

        130kDa: Immature form;

        170kDa: Glycosylated mature form. (PMID:24344333, 24532792, 24705369 and 25233900)

      • Western blot - Anti-ApoER2 antibody [EPR3326] (ab108208)
        Western blot - Anti-ApoER2 antibody [EPR3326] (ab108208)
        All lanes : Anti-ApoER2 antibody [EPR3326] (ab108208) at 1/1000 dilution (unpurified)

        Lane 1 : SH-SY5Y cell lysate
        Lane 2 : C6 cell lysate
        Lane 3 : Neuro-2a cell lysate
        Lane 4 : Human Fetal Brain cell lysate
        Lane 5 : Mouse Fetal Brain cell lysate
        Lane 6 : Rat Fetal Brain cell lysate

        Lysates/proteins at 10 µg per lane.

        Predicted band size: 106 kDa

      • Anti-ApoER2 antibody [EPR3326] (ab108208)
        Anti-ApoER2 antibody [EPR3326] (ab108208)

      References (14)

      Publishing research using ab108208? Please let us know so that we can cite the reference in this datasheet.

      ab108208 has been referenced in 14 publications.

      • Cai J  et al. Genome-scale CRISPR activation screening identifies a role of LRP8 in Sorafenib resistance in Hepatocellular carcinoma. Biochem Biophys Res Commun 526:1170-1176 (2020). PubMed: 32312520
      • Xiang J  et al. Melatonin-induced ApoE expression in mouse astrocytes protects endothelial cells from OGD-R induced injuries. Transl Psychiatry 10:181 (2020). PubMed: 32513932
      • Maire V  et al. LRP8 is overexpressed in estrogen-negative breast cancers and a potential target for these tumors. Cancer Med 8:325-336 (2019). PubMed: 30575334
      • Sasuclark AR  et al. Cell-Type Specific Analysis of Selenium-Related Genes in Brain. Antioxidants (Basel) 8:N/A (2019). PubMed: 31060314
      • Dlugosz P  et al. Differential Action of Reelin on Oligomerization of ApoER2 and VLDL Receptor in HEK293 Cells Assessed by Time-Resolved Anisotropy and Fluorescence Lifetime Imaging Microscopy. Front Mol Neurosci 12:53 (2019). PubMed: 30873003
      View all Publications for this product

      Customer reviews and Q&As

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      Western blot abreview for Anti-ApoER2 antibody [EPR3326]

       Excellent
      Abreviews
      abreview image
      Application
      Western blot
      Sample
      Mouse Tissue lysate - whole (Human THP-1 cell line, various mouse tissues)
      Loading amount
      15 µg
      Specification
      Human THP-1 cell line, various mouse tissues
      Gel Running Conditions
      Reduced Denaturing (4-12% Bis-Tris gel (Novex))
      Blocking step
      Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 22°C
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      Verified customer

      Submitted Nov 02 2012

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