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    apoer2-antibody-epr3326-ab108208.pdf

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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-ApoER2 antibody [EPR3326] (ab108208)

  • Datasheet
Reviews (1) Submit a question References (18)

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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

You may also be interested in

Primary
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Anti-FPR1 antibody (ab113531)
Protein
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Recombinant Human ApoER2 protein (ab152817)
Secondary
Product image
Goat Anti-Rabbit IgG H&L (HRP) (ab205718)

View more associated products

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

  • Form

    Liquid
  • Storage instructions

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

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 40% Glycerol (glycerin, glycerine), 0.05% BSA, 59% PBS
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR3326
  • Isotype

    IgG
  • Research areas

    • Neuroscience
    • Neurology process
    • Neurodegenerative disease
    • Other
    • Neuroscience
    • Neurology process
    • Neurogenesis
    • Signal Transduction
    • Metabolism
    • Lipid metabolism
    • Cardiovascular
    • Lipids / Lipoproteins
    • Lipoproteins/Apolipoproteins
    • Apolipoproteins
    • Metabolism
    • Types of disease
    • Obesity
    • Metabolism
    • Types of disease
    • Cancer

Associated products

  • Alternative Versions

    • Anti-ApoER2 antibody [EPR3326] - BSA and Azide free (ab215274)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
    • Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Positive Controls

    • Rat brain normal tissue lysate - membrane extract (ab29473)
    • Mouse brain tissue lysate - total protein (ab30151)
  • Recombinant Protein

    • Recombinant Human ApoER2 protein (ab152817)

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.
  • Target information above from: UniProt accession Q14114 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 7804 Human
    • Entrez Gene: 16975 Mouse
    • Entrez Gene: 362558 Rat
    • Omim: 602600 Human
    • SwissProt: Q14114 Human
    • SwissProt: Q924X6 Mouse
    • Unigene: 726256 Human
    • Unigene: 442134 Mouse
    • 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)

    Protocols

    • Western blot protocols
    • Immunoprecipitation protocols

    Click here to view the general protocols

    Datasheets and documents

    • Datasheet download

      Download

    References (18)

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

    ab108208 has been referenced in 18 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
    • Kohno T  et al. Reelin-Nrp1 Interaction Regulates Neocortical Dendrite Development in a Context-Specific Manner. J Neurosci 40:8248-8261 (2020). PubMed: 33009002
    • Gao J  et al. Therapeutic IDOL Reduction Ameliorates Amyloidosis and Improves Cognitive Function in APP/PS1 Mice. Mol Cell Biol 40:N/A (2020). PubMed: 31964754
    • Kiroski I  et al. Reelin Improves Cognition and Extends the Lifespan of Mutant Ndel1 Mice with Postnatal CA1 Hippocampus Deterioration. Cereb Cortex 30:4964-4978 (2020). PubMed: 32328622
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    Western blot abreview for Anti-ApoER2 antibody [EPR3326]

    Excellent
    Abreviews
    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
    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 Nov 02 2012

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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