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    apolipoprotein-e-antibody-ep1374y-ab52607.pdf

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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-Apolipoprotein E antibody [EP1374Y] (ab52607)

  • Datasheet
  • SDS
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Western blot - Anti-Apolipoprotein E antibody [EP1374Y] (ab52607)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Apolipoprotein E antibody [EP1374Y] (ab52607)
  • Western blot - Anti-Apolipoprotein E antibody [EP1374Y] (ab52607)
  • Immunocytochemistry/ Immunofluorescence - Anti-Apolipoprotein E antibody [EP1374Y] (ab52607)
  • Western blot - Anti-Apolipoprotein E antibody [EP1374Y] (ab52607)
  • Flow Cytometry (Intracellular) - Anti-Apolipoprotein E antibody [EP1374Y] (ab52607)
  • Immunoprecipitation - Anti-Apolipoprotein E antibody [EP1374Y] (ab52607)
  • Immunoprecipitation - Anti-Apolipoprotein E antibody [EP1374Y] (ab52607)
  • Anti-Apolipoprotein E antibody [EP1374Y] (ab52607)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EP1374Y] to Apolipoprotein E
  • Suitable for: Flow Cyt (Intra), WB, IP, IHC-P, ICC/IF
  • Knockout validated
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Alexa Fluor® 647 Carrier Free HRP

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Overview

  • Product name

    Anti-Apolipoprotein E antibody [EP1374Y]
    See all Apolipoprotein E primary antibodies
  • Description

    Rabbit monoclonal [EP1374Y] to Apolipoprotein E
  • Host species

    Rabbit
  • Tested applications

    Suitable for: Flow Cyt (Intra), WB, IP, IHC-P, ICC/IFmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

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

  • Positive control

    • WB: HEK293 and HepG2 cell lysate, Human liver, cerebellum and serum lysate; Flow Cyt (intra): HepG2 cells; IP: HepG2 whole cell lysate and Human serum; ICC/IF: HepG2 cell lysate; IHC: Astrocytoma tissue.
  • 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.

    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.
  • Storage buffer

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

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EP1374Y
  • Isotype

    IgG
  • Research areas

    • Neuroscience
    • Neurology process
    • Neurodegenerative disease
    • Alzheimer's disease
    • Other
    • Cardiovascular
    • Lipids / Lipoproteins
    • Lipid Metabolism
    • Cholesterol Metabolism
    • Cardiovascular
    • Lipids / Lipoproteins
    • Lipoproteins/Apolipoproteins
    • Neuroscience
    • Neurology process
    • Neurodegenerative disease
    • Other
    • Stem Cells
    • Lineage Markers
    • Endoderm
    • Signal Transduction
    • Metabolism
    • Lipid metabolism
    • Cardiovascular
    • Atherosclerosis
    • Lipid transport
    • Developmental Biology
    • Lineage specification
    • Endoderm
    • Cancer
    • Cancer Metabolism
    • Metabolic signaling pathway
    • Metabolism of lipids and lipoproteins
    • Metabolism
    • Pathways and Processes
    • Metabolic signaling pathways
    • Lipid and lipoprotein metabolism
    • Lipid metabolism
    • Metabolism
    • Pathways and Processes
    • Metabolic signaling pathways
    • Lipid and lipoprotein metabolism
    • Cholesterol Metabolism
    • Metabolism
    • Types of disease
    • Neurodegenerative disease
    • Metabolism
    • Types of disease
    • Cancer
    • Metabolism
    • Types of disease
    • Heart disease
    • Neuroscience
    • Diseases

Associated products

  • Alternative Versions

    • Anti-Apolipoprotein E antibody [EP1374Y] - Low endotoxin, Azide free (ab184845)
    • HRP Anti-Apolipoprotein E antibody [EP1374Y] (ab195855)
    • Alexa Fluor® 647 Anti-Apolipoprotein E antibody [EP1374Y] (ab196194)
    • Alexa Fluor® 488 Anti-Apolipoprotein E antibody [EP1374Y] (ab196463)
    • Anti-Apolipoprotein E antibody [EP1374Y] - BSA and Azide free (ab271843)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Recombinant Protein

    • Recombinant Human Apolipoprotein E (ab55210)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab52607 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
Flow Cyt (Intra)
1/20.
WB (1)
1/1000 - 1/10000. Detects a band of approximately 36 kDa (predicted molecular weight: 36 kDa).
IP
1/20.
IHC-P
1/800. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF (1)
1/50.
Notes
Flow Cyt (Intra)
1/20.
WB
1/1000 - 1/10000. Detects a band of approximately 36 kDa (predicted molecular weight: 36 kDa).
IP
1/20.
IHC-P
1/800. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF
1/50.

Target

  • Function

    Mediates the binding, internalization, and catabolism of lipoprotein particles. It can serve as a ligand for the LDL (apo B/E) receptor and for the specific apo-E receptor (chylomicron remnant) of hepatic tissues.
  • Tissue specificity

    Occurs in all lipoprotein fractions in plasma. It constitutes 10-20% of very low density lipoproteins (VLDL) and 1-2% of high density lipoproteins (HDL). APOE is produced in most organs. Significant quantities are produced in liver, brain, spleen, lung, adrenal, ovary, kidney and muscle.
  • Involvement in disease

    Defects in APOE are a cause of hyperlipoproteinemia type 3 (HLPP3) [MIM:107741]; also known as familial dysbetalipoproteinemia. Individuals with HLPP3 are clinically characterized by xanthomas, yellowish lipid deposits in the palmar crease, or less specific on tendons and on elbows. The disorder rarely manifests before the third decade in men. In women, it is usually expressed only after the menopause. The vast majority of the patients are homozygous for APOE*2 alleles. More severe cases of HLPP3 have also been observed in individuals heterozygous for rare APOE variants. The influence of APOE on lipid levels is often suggested to have major implications for the risk of coronary artery disease (CAD). Individuals carrying the common APOE*4 variant are at higher risk of CAD.
    Genetic variations in APOE are associated with Alzheimer disease type 2 (AD2) [MIM:104310]. It is a late-onset neurodegenerative disorder characterized by progressive dementia, loss of cognitive abilities, and deposition of fibrillar amyloid proteins as intraneuronal neurofibrillary tangles, extracellular amyloid plaques and vascular amyloid deposits. The major constituent of these plaques is the neurotoxic amyloid-beta-APP 40-42 peptide (s), derived proteolytically from the transmembrane precursor protein APP by sequential secretase processing. The cytotoxic C-terminal fragments (CTFs) and the caspase-cleaved products such as C31 derived from APP, are also implicated in neuronal death. Note=The APOE*4 allele is genetically associated with the common late onset familial and sporadic forms of Alzheimer disease. Risk for AD increased from 20% to 90% and mean age at onset decreased from 84 to 68 years with increasing number of APOE*4 alleles in 42 families with late onset AD. Thus APOE*4 gene dose is a major risk factor for late onset AD and, in these families, homozygosity for APOE*4 was virtually sufficient to cause AD by age 80. The mechanism by which APOE*4 participates in pathogenesis is not known.
    Defects in APOE are a cause of sea-blue histiocyte disease (SBHD) [MIM:269600]; also known as sea-blue histiocytosis. This disorder is characterized by splenomegaly, mild thrombocytopenia and, in the bone marrow, numerous histiocytes containing cytoplasmic granules which stain bright blue with the usual hematologic stains. The syndrome is the consequence of an inherited metabolic defect analogous to Gaucher disease and other sphingolipidoses.
    Defects in APOE are a cause of lipoprotein glomerulopathy (LPG) [MIM:611771]. LPG is an uncommon kidney disease characterized by proteinuria, progressive kidney failure, and distinctive lipoprotein thrombi in glomerular capillaries. It mainly affects people of Japanese and Chinese origin. The disorder has rarely been described in Caucasians.
  • Sequence similarities

    Belongs to the apolipoprotein A1/A4/E family.
  • Post-translational
    modifications

    Synthesized with the sialic acid attached by O-glycosidic linkage and is subsequently desialylated in plasma. O-glycosylated with core 1 or possibly core 8 glycans. Thr-307 is a minor glycosylation site compared to Ser-308.
    Glycated in plasma VLDL of normal subjects, and of hyperglycemic diabetic patients at a higher level (2-3 fold).
    Phosphorylation sites are present in the extracelllular medium.
  • Cellular localization

    Secreted.
  • Target information above from: UniProt accession P02649 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 348 Human
    • Omim: 107741 Human
    • SwissProt: P02649 Human
    • Unigene: 654439 Human
    • Alternative names

      • AD2 antibody
      • Apo-E antibody
      • APOE antibody
      • APOE_HUMAN antibody
      • APOEA antibody
      • Apolipoprotein E antibody
      • Apolipoprotein E3 antibody
      • ApolipoproteinE antibody
      • Apoprotein antibody
      • LDLCQ5 antibody
      • LPG antibody
      see all

    Images

    • Western blot - Anti-Apolipoprotein E antibody [EP1374Y] (ab52607)
      Western blot - Anti-Apolipoprotein E antibody [EP1374Y] (ab52607)
      All lanes : Anti-Apolipoprotein E antibody [EP1374Y] (ab52607) at 1/1000 dilution

      Lane 1 : Wild-type HepG2 cell lysate
      Lane 2 : APOE knockout HepG2 cell lysate
      Lane 3 : Human Liver cell lysate
      Lane 4 : Human Kidney cell lysate

      Lysates/proteins at 20 µg per lane.

      Performed under reducing conditions.

      Predicted band size: 36 kDa
      Observed band size: 34 kDa why is the actual band size different from the predicted?



      False colour image of Western blot: Anti-Apolipoprotein E antibody [EP1374Y] staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab52607 was shown to bind specifically to Apolipoprotein E. A band was observed at 34-37 kDa in wild-type HepG2 cell lysates with no signal observed at this size in APOE knockout cell line. To generate this image, wild-type and APOE knockout HepG2 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Apolipoprotein E antibody [EP1374Y] (ab52607)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Apolipoprotein E antibody [EP1374Y] (ab52607)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human astrocytoma tissue sections labeling Apolipoprotein E with purified ab52607 at 1/800 dilution (0.13 µg/mL). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
    • Western blot - Anti-Apolipoprotein E antibody [EP1374Y] (ab52607)
      Western blot - Anti-Apolipoprotein E antibody [EP1374Y] (ab52607)
      All lanes : Anti-Apolipoprotein E antibody [EP1374Y] (ab52607) at 1/10000 dilution (Purified)

      Lane 1 : Human liver lysate
      Lane 2 : Human cerebellum lysate
      Lane 3 : HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysate
      Lane 4 : Human serum lysate

      Secondary
      All lanes : Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

      Predicted band size: 36 kDa

    • Immunocytochemistry/ Immunofluorescence - Anti-Apolipoprotein E antibody [EP1374Y] (ab52607)
      Immunocytochemistry/ Immunofluorescence - Anti-Apolipoprotein E antibody [EP1374Y] (ab52607)
      Immunocytochemistry analysis of HepG2 (Human hepatocellular carcinoma epithelial cell) cells labeling Apolipoprotein E with Purified ab52607 at 1:50 dilution (2.0 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488,ab150077) was used as the secondary antibody at 1:1000 (2 µg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
    • Western blot - Anti-Apolipoprotein E antibody [EP1374Y] (ab52607)
      Western blot - Anti-Apolipoprotein E antibody [EP1374Y] (ab52607)
      All lanes : Anti-Apolipoprotein E antibody [EP1374Y] (ab52607) at 1/1000 dilution (Unpurified)

      Lane 1 : Wild-type HEK293T cell lysate
      Lane 2 : APOE knockout HEK293T cell lysate
      Lane 3 : HepG2 cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

      Predicted band size: 36 kDa
      Observed band size: 36 kDa



      Lanes 1-3: Merged signal (red and green). Green - ab52607 observed at 36 kDa. Red - loading control ab7291 observed at 50 kDa. 

       ab52607 Anti-Apolipoprotein E antibody [EP1374Y] was shown to specifically react with Apolipoprotein E in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266546 (knockout cell lysate ab256838) was used.  Wild-type and Apolipoprotein E knockout samples were subjected to SDS-PAGE.  ab52607 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated at room temperature for 2. 5 hours at 1 in 1000 dilution and 1 in 20000 dilution respectively.  Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

    • Flow Cytometry (Intracellular) - Anti-Apolipoprotein E antibody [EP1374Y] (ab52607)
      Flow Cytometry (Intracellular) - Anti-Apolipoprotein E antibody [EP1374Y] (ab52607)

      Intracellular Flow Cytometry analysis of HepG2 (Human hepatocellular carcinoma epithelial cell) cells labeling Apolipoprotein E (red) with purified ab52607 (Unpurified) at a 1/250 dilution (10ug/mL). Cells were fixed with 4% paraformaldehyde and permeabilized with 90% methanol. A goat anti rabbit IgG (Alexa Fluorr®488) (ab150077) was used as the secondary antibody at a 1/2000 dilution. Black - Rabbit monoclonal IgG (Black) (ab172730). Blue (unlabeled control) - Cell without incubation with primary antibody and secondary antibody (Blue). 

       

       

    • Immunoprecipitation - Anti-Apolipoprotein E antibody [EP1374Y] (ab52607)
      Immunoprecipitation - Anti-Apolipoprotein E antibody [EP1374Y] (ab52607)

      Purified ab52607 at 1/20 dilution (0.5µg) immunoprecipitating Apolipoprotein E in HepG2 whole cell lysate.
      Lane 1 (input): HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysate 10µg
      Lane 2 (+): ab52607 + HepG2 whole cell lysate.
      Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab52607 in HepG2 whole cell lysate.
      VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/5000 dilution) was used for Western blotting.
      Blocking Buffer and concentration: 5% NFDM/TBST.
      Diluting buffer and concentration: 5% NFDM/TBST.
      Observed band size: 36 kDa

    • Immunoprecipitation - Anti-Apolipoprotein E antibody [EP1374Y] (ab52607)
      Immunoprecipitation - Anti-Apolipoprotein E antibody [EP1374Y] (ab52607)

      Purified ab52607 at 1/20 dilution (0.5µg) immunoprecipitating Apolipoprotein E in Human serum.
      Lane 1 (input): Human serum 10µg
      Lane 2 (+): ab52607 + Human serum.
      Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab52607 in Human serum.
      VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/1000 dilution) was used for Western blotting.
      Blocking Buffer and concentration: 5% NFDM/TBST.
      Diluting buffer and concentration: 5% NFDM/TBST.
      Observed band size: 36 kDa

    • Anti-Apolipoprotein E antibody [EP1374Y] (ab52607)
      Anti-Apolipoprotein E antibody [EP1374Y] (ab52607)

    Protocols

    To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (22)

    Publishing research using ab52607? Please let us know so that we can cite the reference in this datasheet.

    ab52607 has been referenced in 22 publications.

    • Lee EG  et al. Redefining transcriptional regulation of the APOE gene and its association with Alzheimer's disease. PLoS One 15:e0227667 (2020). PubMed: 31978088
    • Rauch JN  et al. LRP1 is a master regulator of tau uptake and spread. Nature 580:381-385 (2020). PubMed: 32296178
    • Blanchard JW  et al. Reconstruction of the human blood-brain barrier in vitro reveals a pathogenic mechanism of APOE4 in pericytes. Nat Med 26:952-963 (2020). PubMed: 32514169
    • Wang Y  et al. 27-Hydroxycholesterol Promotes the Transfer of Astrocyte-Derived Cholesterol to Neurons in Co-cultured SH-SY5Y Cells and C6 Cells. Front Cell Dev Biol 8:580599 (2020). PubMed: 33330456
    • Lassen S  et al. Perilipin-2 is critical for efficient lipoprotein and hepatitis C virus particle production. J Cell Sci 132:N/A (2019). PubMed: 30559250
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
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    1-3 of 3 Abreviews

    Immunohistochemistry (Frozen sections) abreview for Anti-Apolipoprotein E antibody [EP1374Y]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Mouse Tissue sections (Brain)
    Permeabilization
    Yes - 0.1 M PBS with 1% Triton X
    Specification
    Brain
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 24°C
    Fixative
    Paraformaldehyde
    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.

    Dr. Ruma Raha-Chowdhury

    Verified customer

    Submitted Feb 05 2016

    Western blot abreview for Anti-Apolipoprotein E antibody [EP1374Y]

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (Mono-Mac 6)
    Loading amount
    50 µg
    Specification
    Mono-Mac 6
    Treatment
    TO-901317
    Gel Running Conditions
    Reduced Denaturing (8%)
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 0.5% · Temperature: RT°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.

    Dr. Elizabeth Kilby

    Verified customer

    Submitted Dec 18 2012

    Immunocytochemistry/ Immunofluorescence abreview for Anti-Apolipoprotein E antibody [EP1374Y]

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (THP1 monocyte-derived macrophages)
    Specification
    THP1 monocyte-derived macrophages
    Fixative
    Paraformaldehyde
    Permeabilization
    Yes - 0.25% Triton X-100 in PBS
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: RT°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.

    Dr. Elizabeth Kilby

    Verified customer

    Submitted Nov 02 2012

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