Invitrogen

APOE Polyclonal Antibody

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This Antibody was verified by Knockout to ensure that the antibody binds to the antigen stated. View Details

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APOE Antibody in Western Blot (WB) — Western blot analysis of APOE was performed by separating 30 µg of human plasma by 10% SDS-PAGE. Proteins were transferred to a membrane and probed with a APOE Polyclonal Antibody (Product # PA5-27088) at a dilution of 1:10000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

APOE Antibody in Western Blot (WB) — Western blot analysis of APOE was performed by loading 20 µg of Hep G2 wild type (Lane 1), Hep G2 treated with Protein Transport Inhibitor(PTI) (1X for 4hr) (Lane 2), Hep G2 CAS9 treated with PTI (1X for 4hr) (Lane 3), Hep G2 APOE KO treated with PTI (1X for 4hr) (Lane 4) whole cell extracts. The blot was probed with Anti-APOE Polyclonal Antibody (Product # PA5-27088) (1:1000 dilution) and Goat anti-Rabbit IgG (H+L), Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036) (1:4000 dilution). Loss of signal upon CRISPR mediated knockout (KO) confirms that antibody is specific to APOE.

APOE Antibody in Western Blot (WB) — Western blot analysis of Apo E using 2.5 µg of A) Conditional medium from human primary preadipocytes and B) Conditional medium from differentiated human primary preadipocytes. Samples were loaded onto a 12% SDS-PAGE gel and probed with an Apo E polyclonal antibody (Product # PA5-27088) at a dilution of 1:2000.

APOE Antibody in Western Blot (WB) — Western blot was performed using Anti-APOE Polyclonal Antibody (Product # PA5-27088) and a 36 kDa band corresponding to APOE was observed in Hep G2 with PTI, Brefeldin treatment and Caco-2 with PTI treatment but was absent in A-431 and Daudi which are reported to be negative. Whole cell extracts (30 µg lysate) of Hep G2 (Lane 1), Hep G2 treated with Protein Transport Inhibitor (PTI; 1x for 4 hours) (Lane 2), Hep G2 treated with Brefeldin (10ng/mL for 90 mins) (Lane 3) Caco-2 (Lane 4), Caco-2 treated with Protein Transport Inhibitor (PTI; 1x for 4 hours) (Lane 5), A-431 (Lane 6), A-431 treated with Protein Transport Inhibitor (PTI; 1x for 4 hours) (Lane 7), Daudi (Lane 8) and Daudi treated with Protein Transport Inhibitor (1x for 4 hours) (Lane 9) were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0322BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:3000 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

APOE Antibody in Western Blot (WB) — Western blot analysis of Apo E using 30 µg HepG2 whole cell lysate. Samples were loaded onto a 12% SDS-PAGE gel and probed with an Apo E polyclonal antibody (Product # PA5-27088) at a dilution of 1:1000.

APOE Antibody in Immunohistochemistry (Paraffin) (IHC (P)) — Immunohistochemistry (Paraffin) analysis of APOE was performed in paraffin-embedded human ovarian cancer tissue using APOE Polyclonal Antibody (Product # PA5-27088) at a dilution of 1:500. Antigen Retrieval: Citrate buffer, pH 6.0, 15 min.

APOE Antibody in Immunoprecipitation (IP) — PA5-27088 immunoprecipitates Apolipoprotein E protein in IP experiments. IP Sample: HepG2 whole cell lysate/extract A: 30 µg whole cell lysate/extract of Apolipoprotein E protein expressing HepG2 cells B: Control with 3 µg of pre-immune rabbit IgG C: Immunoprecipitation of Apolipoprotein E by 3 µg of Apolipoprotein E antibody (Product # PA5-27088) on 10% SDS-PAGE.

APOE Antibody in Immunocytochemistry (ICC/IF) — Immunocytochemistry-Immunofluorescence analysis of APOE was performed in HepG2 cells fixed in 4% paraformaldehyde at RT for 15 min. Green: APOE Polyclonal Antibody (Product # PA5-27088) diluted at 1:500. Blue: Hoechst 33342 staining.

APOE Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescence analysis of APOE was performed using 70% confluent log phase Hep G2 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with APOE Rabbit Polyclonal Antibody (Product # PA5-27088) at 5 µg/mL in 0.1% BSA, incubated at 4 degree Celsius overnight and then with Goat anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32731) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

APOE Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescent analysis of Apo E showing staining in the cytoplasm of THP-1 cells. THP-1 cells were fixed in ice-cold MeOH for 5 min and stained using an Apo E polyclonal antibody (Product # PA5-27088) diluted at 1:500. Blue: Hoechst 33342 staining.

APOE Antibody in Knockout — Antibody specificity was demonstrated by CRISPR-Cas9 mediated knockout of target protein. A loss of signal was observed for target protein in APOE (KO) cell line compared to control cell line using Anti-ApoE Polyclonal Antibody (Product # PA5-27088). {KO}

APOE Antibody in Relative expression — Antibody specificity was demonstrated by detection of differential basal expression of the target across cell lysates tested owing to their inherent genetic constitution. Relative expression of APOE was observed in Hep G2 and Caco-2 as compared to A-431 and Daudi using Anti-APOE Monoclonal Antibody (E6D7) (Product # PA5-27088) in Western Blot. {RE}

APOE Antibody — Fig. 3 SEM analysis of the SPR-chip surface. a Control sample in absence of added ApoE to probe for non-specific binding where the immobilized fibrils on the SPR-chip have been sequentially probed with anti-ApoE antibodies and protein-A conjugated 15 nm gold-beads. b Complete setup where fibrils bound to the SPR-chip have been sequentially probed with ApoE4, anti-ApoE antibodies and protein-A conjugated 15 nm gold-beads. Scale bar is 100 nm

Product Details

Applications

Tested Dilution

Publications

Western Blot (WB)

1:500-1:20000

-

Immunohistochemistry (Paraffin) (IHC (P))

1:100-1:1000

-

Immunocytochemistry (ICC)

-

View 1 publication 1 publication

ELISA (ELISA)

1:1000-1:10000

-

Immunoprecipitation (IP)

1:100-1:500

-

Immunocytochemistry (ICC/IF)

1:100-1:1000

-

Product Specifications

Species Reactivity

Human

Published species

Not Applicable

Host/Isotype

Rabbit / IgG

Class

Polyclonal

Type

Antibody

Immunogen

Synthetic peptide corresponding to a region within amino acids 255 and 317 of Human Apo E

Conjugate

Unconjugated

Form

Liquid

Concentration

0.66 mg/mL

Purification

Antigen affinity chromatography

Storage buffer

PBS, pH 7, with 20% glycerol

Contains

0.025% Proclin 300

Storage conditions

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

RRID

AB_2544564

Product Specific Information

Recommended positive controls: HepG2, Conditional medium from differentiated human primary preadipocyte, human plasma.

Store product as a concentrated solution. Centrifuge briefly prior to opening the vial.

Target Information

Apo E (Apolipoprotein E) plays an important role in the metabolism of lipids in the plasma, and is also is a constituent of various plasma lipoprotein-lipid particles. The protein is synthesized in liver, brain, spleen, and the kidney, and concentrations are high in interstitial fluids, where it plays a role in distributing cholesterol from cells where cholesterol is in excess to cells which require cholesterol. Apo E also plays an important role in the formation of very low density lipoprotein and chylomicrons. The Apo E gene is located on chromosome 19 at position 19q13. 2.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Bioinformatics

Protein Aliases: Apo-E; ApoE4; Apolipoprotein; Apolipoprotein E; apolipoprotein E3; apolipoprotein E4; B2G1; BG; MGC1571

Gene Aliases: AD2; APO-E; APOE; LDLCQ5; LPG

UniProt ID: (Human) P02649

Entrez Gene ID: (Human) 348

response to reactive oxygen species retinoid metabolic process negative regulation of endothelial cell proliferation response to dietary excess triglyceride metabolic process cholesterol catabolic process cellular calcium ion homeostasis receptor-mediated endocytosis cytoskeleton organization G-protein coupled receptor signaling pathway nitric oxide mediated signal transduction synaptic transmission, cholinergic cholesterol metabolic process regulation of gene expression negative regulation of platelet activation positive regulation of cholesterol esterification positive regulation of cholesterol efflux long-chain fatty acid transport protein import virion assembly triglyceride catabolic process cGMP-mediated signaling negative regulation of blood coagulation regulation of axon extension positive regulation of cGMP biosynthetic process neuron projection regeneration regulation of Cdc42 protein signal transduction positive regulation of low-density lipoprotein particle receptor catabolic process cholesterol efflux phospholipid efflux very-low-density lipoprotein particle remodeling low-density lipoprotein particle remodeling high-density lipoprotein particle remodeling high-density lipoprotein particle assembly chylomicron remnant clearance high-density lipoprotein particle clearance very-low-density lipoprotein particle clearance lipoprotein metabolic process lipoprotein biosynthetic process lipoprotein catabolic process vasodilation cholesterol homeostasis negative regulation of MAP kinase activity negative regulation of neuron apoptotic process negative regulation of blood vessel endothelial cell migration reverse cholesterol transport positive regulation by host of viral process negative regulation of cholesterol biosynthetic process positive regulation of lipid biosynthetic process intracellular transport regulation of neuronal synaptic plasticity artery morphogenesis negative regulation of inflammatory response positive regulation of nitric-oxide synthase activity positive regulation of membrane protein ectodomain proteolysis maintenance of location in cell fatty acid homeostasis positive regulation of dendritic spine development negative regulation of canonical Wnt signaling pathway AMPA glutamate receptor clustering NMDA glutamate receptor clustering cellular oxidant detoxification regulation of beta-amyloid clearance negative regulation of neuron death positive regulation of postsynaptic membrane organization negative regulation of presynaptic membrane organization negative regulation of beta-amyloid formation positive regulation of dendritic spine maintenance positive regulation of phospholipid efflux positive regulation of lipid transport across blood brain barrier

Process(es)

beta-amyloid binding lipid transporter activity protein binding phospholipid binding heparin binding lipid binding cholesterol binding antioxidant activity cholesterol transporter activity identical protein binding protein homodimerization activity metal chelating activity tau protein binding low-density lipoprotein particle receptor binding phosphatidylcholine-sterol O-acyltransferase activator activity very-low-density lipoprotein particle receptor binding lipoprotein particle binding