- PA5-27088 - Invitrogen Antibodies APOE antibody

Submitted references Plaque-associated human microglia accumulate lipid droplets in a chimeric model of Alzheimer's disease.

Claes C, Danhash EP, Hasselmann J, Chadarevian JP, Shabestari SK, England WE, Lim TE, Hidalgo JLS, Spitale RC, Davtyan H, Blurton-Jones M
Molecular neurodegeneration 2021 Jul 23;16(1):50

Morphological analysis of Apolipoprotein E binding to Aβ Amyloid using a combination of Surface Plasmon Resonance, Immunogold Labeling and Scanning Electron Microscopy.

Islam T, Gharibyan AL, Lee CC, Olofsson A
BMC biotechnology 2019 Dec 11;19(1):97

Complement-Mediated Regulation of Apolipoprotein E in Cultured Human RPE Cells.

Yang P, Skiba NP, Tewkesbury GM, Treboschi VM, Baciu P, Jaffe GJ
Investigative ophthalmology & visual science 2017 Jun 1;58(7):3073-3085

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: 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.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western Blot using APOE Polyclonal Antibody (Product # PA5-27088). Sample (2.5 µg of whole cell lysate). Lane A: Conditional medium from human primary preadipocyte. Lane B: Conditional medium from differentiated human primary preadipocyte. 12% SDS PAGE. APOE Polyclonal Antibody (Product # PA5-27088) diluted at 1:2,000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: APOE Polyclonal Antibody detects APOE protein by western blot analysis. A. 30 µg HepG2 whole cell lysate/extract.12% SDS-PAGE. APOE Polyclonal Antibody (Product # PA5-27088) dilution: 1:1,000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: 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.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: 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).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Sandwich ELISA detection of recombinant full-length Apolipoprotein E protein using APOE Polyclonal Antibody (Product # PA5-27088) as capture antibody at concentration of 5 µg/mL and a mouse monoclonal anti-Apolipoprotein E antibody as detection antibody at concentration of 1 µg/mL. Mouse IgG antibody (HRP) was diluted at 1:10,000 and used to detect the primary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Sandwich ELISA detection of recombinant full-length human apolipoprotein E using a APOE Monoclonal Antibody as capture antibody at concentration of 5 µg/mL and APOE Polyclonal Antibody (Product # PA5-27088) as detection antibody at concentration of 1 µg/mL. Rabbit IgG antibody (HRP) was diluted at 1:10,000 and used to detect the primary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: 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.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: APOE Polyclonal Antibody [C2C3], C-term detects Apolipoprotein E protein at cytoplasm by immunofluorescent analysis. Sample: THP-1 cells were fixed in ice-cold MeOH for 5 min. Green: Apolipoprotein E protein stained by APOE Polyclonal Antibody [C2C3], C-term (Product # PA5-27088) diluted at 1:500. Blue: Hoechst 33342 staining.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: 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.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: APOE Polyclonal Antibody 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 APOE Polyclonal Antibody (Product # PA5-27088). 10% SDS-PAGE. The immunoprecipitated Apolipoprotein E protein was detected by APOE Polyclonal Antibody (Product # PA5-27088) diluted at 1:1,000. Anti-rabbit IgG (HRP) was used as a secondary reagent.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: 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

PA5-27088

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