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PeproTech recombinant apoe2
Recombinant Apoe2, supplied by PeproTech, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/recombinant+apoe2/pmc12019328__41598_2025_96131_MOESM1_ESM-137-0-5?v=PeproTech
Average 90 stars, based on 1 article reviews

recombinant apoe2 - by Bioz Stars, 2026-07

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Molecular docking and simulation analyses of the interaction between ApoE and ACE2. A-C Predicted binding mode of ACE2 with ApoE2, ApoE3 or ApoE4 (the orange region of the ACE2 protein represents the region that binds to the S1 unit of the spike protein, and amino acids 112 and 158 of the ApoE protein are represented by spheres). D-F The three-dimensional mode and structural representation of the interface residues of the ApoE and ACE2 complexes (dotted green lines indicate hydrogen bonds, and blue and purple lines represent amino acid residues in ACE2 and ApoE proteins, respectively)

Journal: Journal of Translational Medicine

Article Title: ApoE4 associated with severe COVID-19 outcomes via downregulation of ACE2 and imbalanced RAS pathway

doi: 10.1186/s12967-023-03945-7

Figure Lengend Snippet: Molecular docking and simulation analyses of the interaction between ApoE and ACE2. A-C Predicted binding mode of ACE2 with ApoE2, ApoE3 or ApoE4 (the orange region of the ACE2 protein represents the region that binds to the S1 unit of the spike protein, and amino acids 112 and 158 of the ApoE protein are represented by spheres). D-F The three-dimensional mode and structural representation of the interface residues of the ApoE and ACE2 complexes (dotted green lines indicate hydrogen bonds, and blue and purple lines represent amino acid residues in ACE2 and ApoE proteins, respectively)

Article Snippet: The recombinant RBD (SinoBiological, 40592-V05H) and recombinant human ACE2 proteins (SinoBiological, 10108-H02H) were captured with an anti-mIgG Fc Capture (AMC) probe in PBST solution and then incubated with recombinant human ApoE2 (PeproTech, 350-12-500UG), ApoE3 (PeproTech, 350-02-500UG) and ApoE4 (PeproTech, 350-04-500UG) proteins.

Techniques: Binding Assay

ApoE interacts with ACE2 and the spike protein in an isoform-independent manner. A Representative images of individual immunofluorescence staining of ApoE and ACE2 interaction tested by Duolink PLA in HEK-293 T cells after 48 h of cotransfected by Myc-tagged ACE2 and 3 × Flag-tagged ApoE. The red particles (ApoE/ACE2 interaction) represent their interaction. DAPI as a nuclear marker. Scale bar: 5 μm. B Plasmids carrying Myc-tagged ACE2 and 3 × Flag-tagged ApoE (ApoE2, ApoE3 or ApoE4) were transiently cotransfected into HEK-293 T cells. After 48 h of transfection, the cell lysates were immunoprecipitated with anti-Myc antibody and subsequently immunoblotted with anti-Myc and anti-Flag antibodies. The data are presented as the mean ± SD, *P < 0.05, **P < 0.01, ***P < 0.001. C Schematic of the working principles of the BLI assay, which include loading, binding and dissociation. The binding affinities of ApoE and ACE2 were determined through BLI experiments. D The sensor surfaces were immersed in a solution of human ACE2 protein (20 µg/ml), and functionalized sensorgrams were captured upon incubation with human ApoE2, ApoE3, and ApoE4 at 735.3 (blue), 1471 (red), 2941 (light blue), and 5882 nM (green) (binding phase); then, the sensors were immersed in washing buffer (dissociation phase). E The binding affinities of ApoE and the SARS-CoV-2 RBD were determined through BLI experiments. The sensors were immersed in a solution of SARS-CoV-2 RBD (20 µg/ml), and functionalized sensorgrams were captured upon incubation with ApoE2, ApoE3, and ApoE4 at 367.6 (green), 735.3 (yellow), 1471 (light blue), 2941 (brown), and 5882 nM (blue); then, the sensors were immersed in washing buffer (dissociation phase)

Journal: Journal of Translational Medicine

Article Title: ApoE4 associated with severe COVID-19 outcomes via downregulation of ACE2 and imbalanced RAS pathway

doi: 10.1186/s12967-023-03945-7

Figure Lengend Snippet: ApoE interacts with ACE2 and the spike protein in an isoform-independent manner. A Representative images of individual immunofluorescence staining of ApoE and ACE2 interaction tested by Duolink PLA in HEK-293 T cells after 48 h of cotransfected by Myc-tagged ACE2 and 3 × Flag-tagged ApoE. The red particles (ApoE/ACE2 interaction) represent their interaction. DAPI as a nuclear marker. Scale bar: 5 μm. B Plasmids carrying Myc-tagged ACE2 and 3 × Flag-tagged ApoE (ApoE2, ApoE3 or ApoE4) were transiently cotransfected into HEK-293 T cells. After 48 h of transfection, the cell lysates were immunoprecipitated with anti-Myc antibody and subsequently immunoblotted with anti-Myc and anti-Flag antibodies. The data are presented as the mean ± SD, *P < 0.05, **P < 0.01, ***P < 0.001. C Schematic of the working principles of the BLI assay, which include loading, binding and dissociation. The binding affinities of ApoE and ACE2 were determined through BLI experiments. D The sensor surfaces were immersed in a solution of human ACE2 protein (20 µg/ml), and functionalized sensorgrams were captured upon incubation with human ApoE2, ApoE3, and ApoE4 at 735.3 (blue), 1471 (red), 2941 (light blue), and 5882 nM (green) (binding phase); then, the sensors were immersed in washing buffer (dissociation phase). E The binding affinities of ApoE and the SARS-CoV-2 RBD were determined through BLI experiments. The sensors were immersed in a solution of SARS-CoV-2 RBD (20 µg/ml), and functionalized sensorgrams were captured upon incubation with ApoE2, ApoE3, and ApoE4 at 367.6 (green), 735.3 (yellow), 1471 (light blue), 2941 (brown), and 5882 nM (blue); then, the sensors were immersed in washing buffer (dissociation phase)

Techniques: Immunofluorescence, Staining, Marker, Transfection, Immunoprecipitation, Binding Assay, Incubation

ApoE4 downregulates ACE2 protein expression in vitro. A-D ACE2 protein levels in SH-SY5Y, HEK-293 T, A549 and HUVEC cells were examined by western blotting after transfection with 1 µg/ml plasmids expressing Flag, ApoE2-Flag, ApoE3-Flag or ApoE4-Flag for 48 h. The results were normalized to the expression of a-tubulin. The data are expressed as the mean ± SD. E Cells were stained with ACE2 (green) antibody and counterstained with DAPI (blue) after transfection with 1 µg/ml Flag, ApoE2-Flag, ApoE3-Flag or ApoE4-Flag plasmids for 48 h. The data are expressed as the mean ± SD. Statistical differences were evaluated by one-way ANOVA. Scale bars, 20 μm. *P < 0.05, **P < 0.01

Journal: Journal of Translational Medicine

Article Title: ApoE4 associated with severe COVID-19 outcomes via downregulation of ACE2 and imbalanced RAS pathway

doi: 10.1186/s12967-023-03945-7

Figure Lengend Snippet: ApoE4 downregulates ACE2 protein expression in vitro. A-D ACE2 protein levels in SH-SY5Y, HEK-293 T, A549 and HUVEC cells were examined by western blotting after transfection with 1 µg/ml plasmids expressing Flag, ApoE2-Flag, ApoE3-Flag or ApoE4-Flag for 48 h. The results were normalized to the expression of a-tubulin. The data are expressed as the mean ± SD. E Cells were stained with ACE2 (green) antibody and counterstained with DAPI (blue) after transfection with 1 µg/ml Flag, ApoE2-Flag, ApoE3-Flag or ApoE4-Flag plasmids for 48 h. The data are expressed as the mean ± SD. Statistical differences were evaluated by one-way ANOVA. Scale bars, 20 μm. *P < 0.05, **P < 0.01

Techniques: Expressing, In Vitro, Western Blot, Transfection, Staining

ApoE4 downregulates ACE2 protein expression in vivo. A ApoE-TR mice were selected to evaluate the regulatory effect of ApoE4 on ACE2 protein expression. B ACE2 protein levels in the heart, liver, lung, kidney, brain, and bowel (equivalent amounts of total proteins) of ApoE3-TR mice were measured by western blotting, n = 4 mice per group. C-I ACE2 protein levels in the heart, liver, cortex, hippocampus, bowel, kidney, and lung of ApoE2-TR, ApoE3-TR and ApoE4-TR mice were measured by western blotting. The results were normalized to the expression of a-tubulin. n = 6 mice per group. The data are expressed as the mean ± SD. Statistical differences were evaluated by one-way ANOVA. *P < 0.05, **P < 0.01, ***P < 0.001

Journal: Journal of Translational Medicine

Article Title: ApoE4 associated with severe COVID-19 outcomes via downregulation of ACE2 and imbalanced RAS pathway

doi: 10.1186/s12967-023-03945-7

Figure Lengend Snippet: ApoE4 downregulates ACE2 protein expression in vivo. A ApoE-TR mice were selected to evaluate the regulatory effect of ApoE4 on ACE2 protein expression. B ACE2 protein levels in the heart, liver, lung, kidney, brain, and bowel (equivalent amounts of total proteins) of ApoE3-TR mice were measured by western blotting, n = 4 mice per group. C-I ACE2 protein levels in the heart, liver, cortex, hippocampus, bowel, kidney, and lung of ApoE2-TR, ApoE3-TR and ApoE4-TR mice were measured by western blotting. The results were normalized to the expression of a-tubulin. n = 6 mice per group. The data are expressed as the mean ± SD. Statistical differences were evaluated by one-way ANOVA. *P < 0.05, **P < 0.01, ***P < 0.001

Techniques: Expressing, In Vivo, Western Blot

ApoE4 regulates Ang II and Ang 1–7 protein expression in vivo. A-F Ang II and Ang 1–7 proteins level in the cortex, kidney, bowel, liver, heart and lung (equivalent amounts of total proteins) of ApoE2-TR, ApoE3-TR, and ApoE4-TR mice were assessed by ELISA, n = 6 mice per group. G-J MasR protein level in the cortex, kidney, bowel and lung of ApoE2-TR, ApoE3-TR, and ApoE4-TR mice were measured by western blotting, n = 4 mice per group. The data are expressed as the mean ± SD. One-way ANOVA tests were used. *p < 0.05; **p < 0.01

Journal: Journal of Translational Medicine

Article Title: ApoE4 associated with severe COVID-19 outcomes via downregulation of ACE2 and imbalanced RAS pathway

doi: 10.1186/s12967-023-03945-7

Figure Lengend Snippet: ApoE4 regulates Ang II and Ang 1–7 protein expression in vivo. A-F Ang II and Ang 1–7 proteins level in the cortex, kidney, bowel, liver, heart and lung (equivalent amounts of total proteins) of ApoE2-TR, ApoE3-TR, and ApoE4-TR mice were assessed by ELISA, n = 6 mice per group. G-J MasR protein level in the cortex, kidney, bowel and lung of ApoE2-TR, ApoE3-TR, and ApoE4-TR mice were measured by western blotting, n = 4 mice per group. The data are expressed as the mean ± SD. One-way ANOVA tests were used. *p < 0.05; **p < 0.01

Techniques: Expressing, In Vivo, Enzyme-linked Immunosorbent Assay, Western Blot

(A) Protein sequence alignment of human and mouse ApoE (human ApoE3 is shown). The three major isoforms of human ApoE differ by two amino acid residues at 112 and 158 (indicated by red circles): ApoE2 (Cyc112, Cys158), ApoE3 (Cys112, Arg158), and ApoE4 (Arg112, Arg158). Human ApoE contains seven O-linked glycosylation residues (indicated by blue squares), whereas there is only one residue (indicated by green square; corresponding to Thr289 in human ApoE) present in mouse ApoE. (B) Human and mouse brain cortex lysates were analyzed by SDS-PAGE and blots were probed for ApoE immunoreactivity with anti-ApoE antibodies. (C) Human and mouse brain cortex lysates were treated with neuraminidase or buffer alone; protein samples were then analyzed by SDS-PAGE and probed for ApoE immunoactivity using anti-ApoE antibodies.

Journal: Neurobiology of disease

Article Title: Human apolipoprotein E isoforms are differentially sialylated and the sialic acid moiety in ApoE2 attenuates ApoE2-Aβ interaction and Aβ fibrillation

doi: 10.1016/j.nbd.2022.105631

Figure Lengend Snippet: (A) Protein sequence alignment of human and mouse ApoE (human ApoE3 is shown). The three major isoforms of human ApoE differ by two amino acid residues at 112 and 158 (indicated by red circles): ApoE2 (Cyc112, Cys158), ApoE3 (Cys112, Arg158), and ApoE4 (Arg112, Arg158). Human ApoE contains seven O-linked glycosylation residues (indicated by blue squares), whereas there is only one residue (indicated by green square; corresponding to Thr289 in human ApoE) present in mouse ApoE. (B) Human and mouse brain cortex lysates were analyzed by SDS-PAGE and blots were probed for ApoE immunoreactivity with anti-ApoE antibodies. (C) Human and mouse brain cortex lysates were treated with neuraminidase or buffer alone; protein samples were then analyzed by SDS-PAGE and probed for ApoE immunoactivity using anti-ApoE antibodies.

Article Snippet: Production and purification of rhApoE2 Recombinant human ApoE2 was produced in HEK293 cells (FreeStyle 293-F cells, Thermo Fisher Scientific).

Techniques: Sequencing, Glycoproteomics, Residue, SDS Page

$(A) Empty vector (pCMV6-Entry), myc-tagged pCMV6-human ApoE2, pCMV6-human ApoE3, or pCMV6-human ApoE4 constructs were transfected into human SH-SY5Y cells. Cell lysates and culture media fractions were harvested and probed for ApoE immunoreactivity using anti-myc antibodies. Pull-down assay was performed with myc-trap on culture media fractions. Results are shown as the intensity ratio of sialylated_ApoE versus nonsialylated_ApoE and presented as mean ± standard deviation. ****p < 0.0001. (B) Myc-tagged pCMV6-human ApoE3 construct was transfected into SH-SY5Y cells. Cell lysates and culture media were harvested and treated with endoglycosidases (PNGase F, Endo H, O-glycosidase, and neuraminidase). The resulting samples were analyzed for ApoE immunoreactivity using anti-myc antibodies. (C) Myc-tagged pCMV6-human ApoE2/ApoE3/ApoE4 constructs were transfected into SH-SY5Y cells. Cell lysates and culture media were harvested, treated with neuraminidase, and probed for ApoE immunoreactivity using anti-myc antibodies.$

Journal: Neurobiology of disease

Article Title: Human apolipoprotein E isoforms are differentially sialylated and the sialic acid moiety in ApoE2 attenuates ApoE2-Aβ interaction and Aβ fibrillation

doi: 10.1016/j.nbd.2022.105631

Figure Lengend Snippet: (A) Empty vector (pCMV6-Entry), myc-tagged pCMV6-human ApoE2, pCMV6-human ApoE3, or pCMV6-human ApoE4 constructs were transfected into human SH-SY5Y cells. Cell lysates and culture media fractions were harvested and probed for ApoE immunoreactivity using anti-myc antibodies. Pull-down assay was performed with myc-trap on culture media fractions. Results are shown as the intensity ratio of sialylated_ApoE versus nonsialylated_ApoE and presented as mean ± standard deviation. ****p < 0.0001. (B) Myc-tagged pCMV6-human ApoE3 construct was transfected into SH-SY5Y cells. Cell lysates and culture media were harvested and treated with endoglycosidases (PNGase F, Endo H, O-glycosidase, and neuraminidase). The resulting samples were analyzed for ApoE immunoreactivity using anti-myc antibodies. (C) Myc-tagged pCMV6-human ApoE2/ApoE3/ApoE4 constructs were transfected into SH-SY5Y cells. Cell lysates and culture media were harvested, treated with neuraminidase, and probed for ApoE immunoreactivity using anti-myc antibodies.

Article Snippet: Production and purification of rhApoE2 Recombinant human ApoE2 was produced in HEK293 cells (FreeStyle 293-F cells, Thermo Fisher Scientific).

Techniques: Plasmid Preparation, Construct, Transfection, Pull Down Assay, Standard Deviation

(A) The purity of recombinant human ApoE2 derived from 293-F cells was analyzed by SDS-PAGE. (B) Sialylation profiles of ApoE2 protein derived from human brain tissue lysates, E. coli, or 293-F cells were comparatively analyzed by immunoblotting. Results are shown as the intensity ratio of sialylated_ApoE2 versus nonsialylated_ApoE2 expressed in human brain tissues of ApoE ε2/ε2 homozygotes in comparison with ApoE2 derived from 293-F cells and presented as mean ± standard deviation. (C) Deglycosylation analyses were performed on rhApoE2 derived from 293-F cells using specific endoglycosidase: PNGase F, Endo H, O-glycosidase, and neuraminidase. The resulting samples were analyzed by immunoblotting using anti-ApoE antibodies.

Journal: Neurobiology of disease

Article Title: Human apolipoprotein E isoforms are differentially sialylated and the sialic acid moiety in ApoE2 attenuates ApoE2-Aβ interaction and Aβ fibrillation

doi: 10.1016/j.nbd.2022.105631

Figure Lengend Snippet: (A) The purity of recombinant human ApoE2 derived from 293-F cells was analyzed by SDS-PAGE. (B) Sialylation profiles of ApoE2 protein derived from human brain tissue lysates, E. coli, or 293-F cells were comparatively analyzed by immunoblotting. Results are shown as the intensity ratio of sialylated_ApoE2 versus nonsialylated_ApoE2 expressed in human brain tissues of ApoE ε2/ε2 homozygotes in comparison with ApoE2 derived from 293-F cells and presented as mean ± standard deviation. (C) Deglycosylation analyses were performed on rhApoE2 derived from 293-F cells using specific endoglycosidase: PNGase F, Endo H, O-glycosidase, and neuraminidase. The resulting samples were analyzed by immunoblotting using anti-ApoE antibodies.

Article Snippet: Production and purification of rhApoE2 Recombinant human ApoE2 was produced in HEK293 cells (FreeStyle 293-F cells, Thermo Fisher Scientific).

Techniques: Recombinant, Derivative Assay, SDS Page, Western Blot, Comparison, Standard Deviation

(A-B) Following a 2 hr at 37 °C incubation of rhApoE2 derived from E. coli or 293-F cells with Aβ peptides, the rhApoE2-Aβ40 (A) and rhApoE2-Aβ42 (B) complexes were analyzed by SDS-PAGE and probed for Aβ and ApoE immunoreactivities. (C) Sialyalted_rhApoE2 (derived from 293-F cells) vs desialylated_rhApoE2 (prepared from rhApoE2 derived from 293-F cells and treated with neuraminidase) were confirmed by immunoblotting. (D) Sialylated_rhApoE2-Aβ40 and desialylated_rhApoE2-Aβ40 complexes were immunoprecipitated using anti-human monoclonal Aβ antibody (6e10). The elutes were probed for ApoE immunoreactivity using goat polyclonal ApoE antibody. (E-G) The interactions of sialylated_rhApoE2 versus desialyated_rhApoE2 with Aβ fragments: (E) Aβ1-16, (F) Aβ12-28, and (G) Aβ25-35, were analyzed by SDS-PAGE and probed for Aβ and ApoE immunoreactivities using specific antibodies as indicated. Quantitative results are shown as relative band intensity and presented as mean ± standard deviation. Each close circle and square in each graph is representative of one independent experiment (n=9 for Aβ1-16, n=7 for Aβ12-28, and n=6 for Aβ25-35). **p < 0.01. (H) The residues indicated in red represent the sequence region in Aβ that preferentially binds desialyated_rhApoE2. (I) Thioflavin T Aβ42 aggregation assay was performed to analyze Aβ42 fibril formation influenced by sialylated_rhApoE2 in comparison with desialylated_rhApoE2. The graph is generated from one experiment but is representative of n=4 independent repeats of the same assay. * p < 0.05, **p < 0.01, and ***p < 0.001. (J) Data presented in this figure indicate that the sialic acid modification in human ApoE protein may serve as a key modulator of Aβ fibrillization. The removal of sialic acid increases ApoE binding affinity for Aβ17-24 residues and such an interaction may promote Aβ fibrillation. The presence of sialic acid attenuates ApoE binding with Aβ17-24 and reduces Aβ fibrillation.

Journal: Neurobiology of disease

Article Title: Human apolipoprotein E isoforms are differentially sialylated and the sialic acid moiety in ApoE2 attenuates ApoE2-Aβ interaction and Aβ fibrillation

doi: 10.1016/j.nbd.2022.105631

Figure Lengend Snippet: (A-B) Following a 2 hr at 37 °C incubation of rhApoE2 derived from E. coli or 293-F cells with Aβ peptides, the rhApoE2-Aβ40 (A) and rhApoE2-Aβ42 (B) complexes were analyzed by SDS-PAGE and probed for Aβ and ApoE immunoreactivities. (C) Sialyalted_rhApoE2 (derived from 293-F cells) vs desialylated_rhApoE2 (prepared from rhApoE2 derived from 293-F cells and treated with neuraminidase) were confirmed by immunoblotting. (D) Sialylated_rhApoE2-Aβ40 and desialylated_rhApoE2-Aβ40 complexes were immunoprecipitated using anti-human monoclonal Aβ antibody (6e10). The elutes were probed for ApoE immunoreactivity using goat polyclonal ApoE antibody. (E-G) The interactions of sialylated_rhApoE2 versus desialyated_rhApoE2 with Aβ fragments: (E) Aβ1-16, (F) Aβ12-28, and (G) Aβ25-35, were analyzed by SDS-PAGE and probed for Aβ and ApoE immunoreactivities using specific antibodies as indicated. Quantitative results are shown as relative band intensity and presented as mean ± standard deviation. Each close circle and square in each graph is representative of one independent experiment (n=9 for Aβ1-16, n=7 for Aβ12-28, and n=6 for Aβ25-35). **p < 0.01. (H) The residues indicated in red represent the sequence region in Aβ that preferentially binds desialyated_rhApoE2. (I) Thioflavin T Aβ42 aggregation assay was performed to analyze Aβ42 fibril formation influenced by sialylated_rhApoE2 in comparison with desialylated_rhApoE2. The graph is generated from one experiment but is representative of n=4 independent repeats of the same assay. * p < 0.05, **p < 0.01, and ***p < 0.001. (J) Data presented in this figure indicate that the sialic acid modification in human ApoE protein may serve as a key modulator of Aβ fibrillization. The removal of sialic acid increases ApoE binding affinity for Aβ17-24 residues and such an interaction may promote Aβ fibrillation. The presence of sialic acid attenuates ApoE binding with Aβ17-24 and reduces Aβ fibrillation.

Article Snippet: Production and purification of rhApoE2 Recombinant human ApoE2 was produced in HEK293 cells (FreeStyle 293-F cells, Thermo Fisher Scientific).

Techniques: Incubation, Derivative Assay, SDS Page, Western Blot, Immunoprecipitation, Standard Deviation, Sequencing, Comparison, Generated, Modification, Binding Assay

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