Invitrogen

APOE Polyclonal Antibody

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问题与答案

问题与答案

问题与答案

抗体检测数据 (7)
高级验证 (2)

APOE Antibody in Immunocytochemistry (ICC/IF)

APOE Antibody in Immunocytochemistry (ICC/IF)

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APOE Antibody (PA5-95232) in ICC/IF

Immunofluorescence analysis of APOE was performed using 70% confluent log phase Hep G2 and A-431 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 45 minutes at room temperature. The cells were labeled with APOE Polyclonal Antibody (Product # PA5-95232) at 5 µg/mL in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A... View More

APOE Antibody in Immunocytochemistry (ICC/IF)

APOE Antibody in Immunohistochemistry (Paraffin) (IHC (P))

APOE Antibody in Immunohistochemistry (Paraffin) (IHC (P))

APOE Antibody in Western Blot (WB)

APOE Antibody in Western Blot (WB)

APOE Antibody in Flow Cytometry (Flow)

APOE Antibody in ELISA (ELISA)

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APOE Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescence analysis of APOE was performed using 70% confluent log phase Hep G2 and A-431 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 45 minutes at room temperature. The cells were labeled with APOE Polyclonal Antibody (Product # PA5-95232) at 5 µg/mL in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32790, 1:2000 dilution), 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 dilution). Panel d represents the merged image showing cytoplasmic localization in Hep G2 but not in A-431, which is reported to be negative for APOE expression (Panel e). Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

APOE Antibody in Immunohistochemistry (Paraffin) (IHC (P)) — Immunohistochemistry (Paraffin) analysis of APOE in paraffin-embedded section of human placenta tissue using APOE Polyclonal Antibody (Product # PA5-95232). Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with the primary antibody at a 5 µg/mL dilution overnight at 4°C. Cy3 conjugated goat anti-rabbit IgG was used as secondary antibody at 1:500 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

APOE Antibody in Immunohistochemistry (Paraffin) (IHC (P)) — Immunohistochemistry analysis of APOE in paraffin-embedded human placenta tissue. Antigen retrieval was performed on the tissue using citrate buffer (pH 6, 20 min) and blocked with 10% goat serum. Samples were incubated with APOE polyclonal antibody (Product # PA5-95232) at a 1 µg/mL dilution, followed by biotinylated goat anti-rabbit IgG (30 min, 37°C), and developed with Strepavidin-Biotin-Complex and DAB.

APOE Antibody in Western Blot (WB) — Western blot was performed using APOE Polyclonal Antibody (Product # PA5-95232) and a 38 kDa band corresponding to APOE was observed only in HepG2 cell line as compared to A-431 and IMR-32 cell lines which are reported to be negative for APOE expression. Whole cell extracts (30 µg lysate) of Hep G2 (Lane 1), A-431 (Lane 2), IMR-32 (Lane 3) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0321BOX), 10 well. Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23002) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (0.5 µg/mL) and detected by chemiluminescence with Goat anti-Rabbit IgG (Heavy Chain) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:20,000 dilution) using the iBright™ FL1500 Imaging System (Product # A44115). Chemiluminescent detection was performed using SuperSignal™ West Pico PLUS Chemiluminescent Substrate (Product # 34580).

APOE Antibody in Western Blot (WB) — Western blot analysis of APOE using APOE Polyclonal Antibody (Product # PA5-95232). Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel)/90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 µg of sample under reducing conditions. Lane 1: human placenta tissue lysates. Lane 2: human HepG2 whole cell lysates. After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with APOE Polyclonal Antibody at 0.5 µg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5,000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit with Tanon 5200 system. A specific band was detected for APOE at approximately 36 kDa. The expected band size for APOE is at 36 kDa.

APOE Antibody in Flow Cytometry (Flow) — Flow cytometry analysis of APOE in HepG2 cells using APOE Polyclonal Antibody (Product # PA5-95232), shown in overlay histogram (blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum, and incubated with the primary antibody (1 μg/1x10^6 cells) for 30 min at 20°C. DyLight 488 conjugated goat anti-rabbit IgG (5-10 µg/1x10^6 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 µg/1x10^6) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.

APOE Antibody in ELISA (ELISA) — Sandwich ELISA analysis of recombinant human Apolipoprotein E/APOE protein. A standard curve was generated using APOE Polyclonal Antibody (Product # PA5-95232).

APOE Antibody — Antibody specificity was demonstrated by the detection of differential basal expression of the target across the cell lines tested, owing to their inherent genetic constitution. APOE expression was found in HepG2 but not in A-431 or IMR-32 cell lines, which have been reported to be negative for APOE using APOE Polyclonal Antibody (Product # PA5-95232) in Western Blot. {RE}

APOE Antibody — Antibody specificity was demonstrated by the detection of differential basal expression of the target across cell models owing to their inherent genetic constitution. Immunofluorescence analysis using APOE Polyclonal Antibody (Product # PA5-95232) shows cytoplasmic staining only in HepG2 when compared to A-431, which is reported to be negative for APOE expression. {RE}

Immunogen image

APOE Polyclonal Antibody

Immunogen Image APOE Polyclonal Antibody

产品信息

PA5-95232

应用

建议稀释比

已发表文章

免疫印迹 (WB)

0.1-0.5 µg/mL

-

免疫组化（石蜡） (IHC (P))

0.5-1 µg/mL

-

免疫细胞化学 (ICC/IF)

5 µg/mL

-

流式细胞分析 (Flow)

1-3 µg/1x10^6 cells

-

酶联免疫吸附实验 (ELISA)

1-5 µg/mL

-

产品规格

种属反应

Human

宿主/亚型

Rabbit / IgG

分类

Polyclonal

类型

Antibody

抗原

E.coli-derived human Apolipoprotein E recombinant protein (Position: K19-H317).

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偶联物

Unconjugated Unconjugated Unconjugated

形式

Lyophilized

浓度

500 µg/mL

纯化类型

Affinity chromatography

保存液

PBS with 5mg BSA

内含物

0.05mg sodium azide

保存条件

-20°C

运输条件

Ambient (domestic); Wet ice (international)

产品详细信息

Human Apolipoprotein E shares 73% and 72% amino acid (aa) sequence identity with mouse and rat Apolipoprotein E, respectively.

Reconstitute with 0.2 mL of distilled water to yield a concentration of 500 µg/mL.

Positive Control - WB: human placenta tissue, human HepG2 whole cell. IHC: Human Placenta tissue. Flow: HepG2 cell.|Store at -20°C for one year from date of receipt. After reconstitution, at 4°C for one month. It can also be aliquotted and stored frozen at -20°C for six months. Avoid repeated freeze-thaw cycles.

靶标信息

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.

仅用于科研。不用于诊断过程。未经明确授权不得转售。

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引用该产品

生物信息学

蛋白别名: Apo-E; ApoE4; Apolipoprotein; Apolipoprotein E; apolipoprotein E3; apolipoprotein E4; Apoprotein; B2G1; BG; MGC1571

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基因别名: AD2; APO-E; APOE; LDLCQ5; LPG

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UniProt ID: (Human) P02649

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Entrez Gene ID: (Human) 348

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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 apolipoprotein

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