Human Apolipoprotein E/ApoE Antibody AF4144: R&D Systems

Human Apolipoprotein E/ApoE Antibody

(1 citations)   
  • Species Reactivity
    Human
  • Specificity
    Detects human Apolipoprotein E/ApoE in direct ELISAs and Western blots.
  • Source
    Polyclonal Goat IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    E. coli-derived recombinant human Apolipoprotein E3/ApoE3
    Lys19-His317
    Accession # P02649
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    0.5-1 µg/mL
    See below
  • Simple Western
    10 µg/mL
    See below
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Data Examples
Detection of Human Apolipoprotein E/ApoE by Western Blot. Western blot shows lysates of human plasma. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human Apolipoprotein E/ApoE Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4144) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF019).
A specific band was detected for Apolipoprotein E/ApoE at approximately 38 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 8.
Detection of Human Apolipoprotein E/
ApoE by Western Blot.
Western blot shows conditioned media from SK‑Mel‑28 human malignant melanoma cell line, HepG2 human hepatocellular carcinoma cell line, and human peripheral blood mononuclear cells (PBMC). PVDF membrane was probed with 0.5 µg/mL of Goat Anti-Human Apolipoprotein E/ApoE Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4144) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). A specific band was detected for Apolipoprotein E/ApoE at approximately 38 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Detection of Human Apolipoprotein E/
ApoE by Simple WesternTM.
Simple Western lane view shows human plasma, loaded at 1:10 dilution. A specific band was detected for Apolipoprotein E/ApoE at approximately 39 kDa (as indicated) using 10 µg/mL of Goat Anti-Human Apolipoprotein E/ApoE Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4144) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Preparation and Storage
  • Reconstitution
    Reconstitute at 0.2 mg/mL in sterile PBS.
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • Stability & Storage
    Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
    • 12 months from date of receipt, -20 to -70 °C as supplied.
    • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
    • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: Apolipoprotein E/ApoE

ApoE is a major protein component of serum LDL, VLDL, HDL, and chylomicrons. It is produced predominantly by hepatocytes, macrophages, and non-neuronal cells in the CNS. ApoE-containing particles transport triglycerides and cholesterol to peripheral tissues for cellular uptake and catabolism (1‑4). Mature human ApoE is a 37 kDa glycoprotein that consists of an N-terminal domain composed of four bundled alpha -helices, plus a hinge region and an extended alpha -helical C-terminal domain (2, 5). Its amphipathic nature and flexible structure enables it to adopt dramatically different conformations upon lipid association (2). ApoE is monomeric in lipid particles, although it forms oligomers when lipid-free (6). ApoE3 is the most abundant of the three common alleles in human; ApoE2 and ApoE4 differ by single aa substitutions (1). Mature human ApoE shares 71% aa sequence identity with mouse and rat ApoE. LDL receptor family proteins preferentially bind and internalize the lipid-bound form of ApoE with the exception of VLDLR which also efficiently internalizes lipid-free ApoE (7, 8). Lipoprotein uptake is facilitated by the initial binding of ApoE to cell surface heparan sulfate proteoglycans (HSPG) (9). Receptor/HSPG binding and lipid interactions primarily involve the N- and C-terminal regions of ApoE, respectively (2). Recycled lipid-free ApoE is formed into HDL particles through interactions with the lipid transporter ABCA1 (10). High cellular sterol content activates the nuclear hormone receptor LXR which promotes increased ApoE synthesis and increased sterol efflux, while low sterol content induces LDL R expression with increased sterol uptake and decreased ApoE production (11). ApoE3 dampens the TNF-alpha induced inflammatory response in vascular endothelial cells (12). In the CNS, ApoE blocks production of the amyloid A beta peptide by inhibiting the gamma -secretase cleavage of APP (13). It also complexes with A beta and promotes A beta internalization via LRP2 (14, 15).

  • References:
    1. Martins, I.J. et al. (2006) Mol. Pschiatry 11:721.
    2. Hatters, D.M. et al. (2006) Trends Biochem. Sci. 31:445.
    3. Heeren, J. et al. (2006) Arterioscler. Thromb. Vasc. Biol. 26:442.
    4. Mahley, R.W. et al. (1984) J. Lipid. Res. 25:1277.
    5. Zannis, V.I. et al. (1984) J. Biol. Chem. 259:5495.
    6. Perugini, M.A. et al. (2000) J. Biol. Chem. 275:36758.
    7. Ruiz, J. et al. (2005) J. Lipid Res. 46:1721.
    8. Chroni, A. et al. (2005) Biochemistry 44:13132.
    9. Futamura, M. et al. (2005) J. Biol. Chem. 280:5414.
    10. Krimbou, L. et al. (2004) J. Lipid. Res. 45:839.
    11. Lucic, D. et al. (2007) J. Lipid Res. 48:366.
    12. Mullick, A.E. et al. (2007) Arterioscler. Thromb. Vasc. Biol. 27:339.
    13. Irizarry, M.C. et al. (2004) J. Neurochem. 90:1132.
    14. Naslund, J. et al. (1995) Neuron 15:219.
    15. Zerbinatti, C.V. et al. (2006) J. Biol. Chem. 281:36180.
  • Entrez Gene IDs:
    348 (Human); 11816 (Mouse); 25728 (Rat)
  • Alternate Names:
    APOE; Apo-E; Apolipoprotein E; apolipoprotein E3; late onset); MGC1571
Related Research Areas
Citations:

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

1 Citations: Showing 1 - 1

  1. MicroRNA-27a regulates lipid metabolism and inhibits hepatitis C virus replication in human hepatoma cells.
    Authors: Shirasaki T, Honda M, Shimakami T, Horii R, Yamashita T, Sakai Y, Sakai A, Okada H, Watanabe R, Murakami S, Yi M, Lemon S, Kaneko S
    J Virol, 2013;87(9):5270-86.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
Isotype Controls
Description Application Cat# Citations Images  

Normal Goat IgG Control

Ctrl AB-108-C 191  
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Secondary Antibodies
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Goat IgG HRP-conjugated Antibody

WB, Simple Western HAF109 19  
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Goat IgG HRP-conjugated Antibody

WB HAF017 15  
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Goat IgG (H+L) PE-conjugated Antibody

Flow F0107 4  
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Donkey Anti-Goat IgG NorthernLights™ NL557-conjugated Antibody

Flow, IHC, ICC NL001 8
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Donkey Anti-Goat IgG NorthernLights™ NL493-conjugated Antibody

Flow, IHC, ICC NL003 5
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Goat IgG (H+L) APC-conjugated Antibody

Flow F0108 6  
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Goat IgG Horseradish Peroxidase-conjugated Antibody

WB HAF019 6  
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Donkey Anti-Goat IgG Antibody

WB AF109 6
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Donkey Anti-Goat IgG Biotinylated Antibody

WB BAF109 3
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Goat IgG VisUCyte HRP Polymer

IHC VC004  
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Goat IgG (H+L) Fluorescein-conjugated Antibody

Flow F0109 2  
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Donkey Anti-Goat IgG NorthernLights™ NL637-conjugated Antibody

Flow, IHC, ICC NL002
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Chicken Anti-Goat IgG Biotinylated Antibody

WB BAF019
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Rabbit Anti-Goat IgG (H+L) Affinity Purified PAb, X Absorbed

WB, ELISA R-401-C-ABS
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Rabbit Anti-Goat IgG Biotinylated Antibody

WB BAF017
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Donkey Anti-Goat IgG (H+L) PerCP-conjugated Antibody

Flow F0124
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