mouse aβ 1 40 elisa kit  (Cusabio)

Journal: Frontiers in Neuroscience

Article Title: 8-weeks aerobic exercise ameliorates cognitive deficit and mitigates ferroptosis triggered by iron overload in the prefrontal cortex of APP Swe / PSEN 1dE9 mice through Xc − /GPx4 pathway

doi: 10.3389/fnins.2024.1453582

Figure Lengend Snippet: Reduction of Aβ 42 deposition in APP/PS1 mice following 8 weeks of aerobic exercise. Western blot protein signals of the PFC in each group (A) . Relative protein expression levels of APP (B) and BACE1 (C) in the PFC of each group as detected by WB. ELISA detection of Aβ 40 in the PFC of each group (D) . Statistical results of the area percentage of Aβ 42 antigen-positive regions in IHC staining of the PFC (E) , and tissue morphology images at 40× (upper) and 500× (lower) magnification (F) . In cases of significant interaction effect, “&” indicates a significant genotype simple effect between AD-E and WT-E with p ≤ 0.05. In (F) , hematoxylin stains nuclei blue, and DAB reveals Aβ 42 antigen-positive expression in brown.

Article Snippet: The Aβ 40 expression in the PFC tissue of mice was determined using mouse Aβ 1–40 sandwich enzyme linked immunosorbent assay (ELISA) kit (Elabscience, E-EL-M3009).

Techniques: Western Blot, Expressing, Enzyme-linked Immunosorbent Assay, Immunohistochemistry

Journal: International Journal of Molecular Sciences

Article Title: Inhibition of Amyloid-β (Aβ)-Induced Cognitive Impairment and Neuroinflammation in CHI3L1 Knockout Mice through Downregulation of ERK-PTX3 Pathway

doi: 10.3390/ijms25105550

Figure Lengend Snippet: CHI3L1 related to AD. Human serum analysis of Aβ 1–40 , Aβ 1–42 , Tau, and CHI3L1 in patients with AD and healthy controls using ELISA ( n = 25). The ( A ) serum levels and ( B ) ROC curves of Aβ 1–40 , Aβ 1–42 , Tau, and CHI3L1. ( C ) Spearman correlation test results between CHI3L1 and Aβ 1–40 , Aβ 1–42 , or Tau. Each value is mean ± S.E.M. from 25 samples. *, Significantly different from control group ( p < 0.05). ***, Significantly different from control group ( p < 0.001). n.s., not significant.

Article Snippet: Aβ 1–42 and Aβ 1–40 levels were determined using each specific mouse Aβ 1–42 enzyme-linked immunosorbent assay (ELISA) Kit and mouse Aβ 1–40 ELISA Kit purchased from CUSABIO (Houston, TX, USA) following the manufacturer’s protocol.

Techniques: Enzyme-linked Immunosorbent Assay, Control

Journal: Bioactive Materials

Article Title: Significance of native PLGA nanoparticles in the treatment of Alzheimer's disease pathology

doi: 10.1016/j.bioactmat.2022.05.030

Figure Lengend Snippet: Characterization of spontaneous Aβ 1-42 aggregation A; ThT assay revealing the dose-dependent (1–20 μM) spontaneous aggregation curves of Aβ 1-42 over 24h. Note the increased aggregation of Aβ 1-42 which reaches a plateau as indicated by ThT levels. B; ThT-stained fluorescence images depicting a dose-dependent (1, 5, 10 and 20 μM) increase in Aβ 1-42 aggregates after 24h. C; Time-dependent TEM micrographs showing the aggregation of Aβ 1-42 leading to the formation of matured fibers over 24h. D & E; DLS data revealing the hydrodynamic radius of monomeric and aggregated Aβ 1-42 . F; Zeta potential measurement for Aβ 1-42 displaying surface charge of −32mV. G; Native PAGE of Aβ 1-42 aggregates showing the presence of higher ordered fibrillar entities after 24h. H; CD spectra for Aβ 1-42 monomers as well as aggregates confirming the formation of beta-sheet following aggregation of the peptide. I; FTIR secondary derivative spectra showing structural conformation Aβ 1-42 after aggregation.

Article Snippet: Dulbecco's modified Eagle's medium (DMEM), neurobasal medium, Hanks' balanced salt solution (HBSS), fetal bovine serum (FBS), Alexa Fluor 488/594 conjugated secondary antibodies, ProLong Gold anti-fade reagent, NuPAGE 4–12% Bis-Tris gels and ELISA kits for the detection of mouse and human Aβ 1–40 and Aβ 1–42 were purchased from R&D systems (Minneapolis, MN, USA).

Techniques: ThT Assay, Staining, Fluorescence, Zeta Potential Analyzer, Clear Native PAGE, Circular Dichroism

Journal: Bioactive Materials

Article Title: Significance of native PLGA nanoparticles in the treatment of Alzheimer's disease pathology

doi: 10.1016/j.bioactmat.2022.05.030

Figure Lengend Snippet: Attenuation of spontaneous Aβ 1-42 aggregation by PLGA A; Graphical representation showing the polymerization of poly l -lactic acid and poly l -glycolic acid to form PLGA nanoparticles. B; TEM images showing the spheroidal nature of PLGA nanoparticles with an average diameter of ∼100 nm. C: DLS histogram of the PLGA nanoparticles displaying the diameter size of ∼100 nm. D; Zeta potential measurement for PLGA nanoparticles revealing the surface charge of ∼8 mV. E & F; PLGA dose-dependently (1–50 μM) attenuates spontaneous aggregation of 10 μM Aβ 1-42 as revealed by ThT fluorescence assay over the 24h reaction period. G; ThT-stained fluorescence images depicting attenuation of 10 μM Aβ 1-42 aggregation over 24h in the absence and presence of 50 μM PLGA. H; TEM micrograph showing direct association of PLGA nanoparticles with Aβ 1-42 fibers. I & J; DLS analysis revealing hydrodynamic radius of 10 μM Aβ 1-42 in the absence (I) and presence of 50 μM PLGA (J). Note the decrease in the hydrodynamic radius of Aβ 1-42 fibers in the presence of PLGA nanoparticles. K; Native PAGE showing the loss of higher-ordered Aβ aggregates in the presence of PLGA and the corresponding filter-trap assay (in the inset) revealing decreased formation of Aβ 1-42 fibers in the presence of PLGA as detected by fibril-specific OC antibody.

Article Snippet: Dulbecco's modified Eagle's medium (DMEM), neurobasal medium, Hanks' balanced salt solution (HBSS), fetal bovine serum (FBS), Alexa Fluor 488/594 conjugated secondary antibodies, ProLong Gold anti-fade reagent, NuPAGE 4–12% Bis-Tris gels and ELISA kits for the detection of mouse and human Aβ 1–40 and Aβ 1–42 were purchased from R&D systems (Minneapolis, MN, USA).

Techniques: Zeta Potential Analyzer, Fluorescence, Staining, Clear Native PAGE, TRAP Assay

Journal: Bioactive Materials

Article Title: Significance of native PLGA nanoparticles in the treatment of Alzheimer's disease pathology

doi: 10.1016/j.bioactmat.2022.05.030

Figure Lengend Snippet: Characterization of molecular interactions between Aβ 1-42 and PLGA A; CD spectra showing decreased beta-sheet formation following incubation of 10 μM Aβ 1-42 in the presence of 50 μM PLGA nanoparticles. B; FTIR secondary derivative spectra of the aggregating Aβ 1-42 samples in the presence ( red) and absence (black) of PLGA nanoparticles. Note the dominance of beta-rich signals after aggregation of Aβ 1-42 and the occurrence of 3 10 α-helix in the presence of PLGA. C; ThT kinetic assays showing that PLGA with 50:50 resomer from Phosphorex (green) and Sigma (red), but not PLGA with 75:25 resomer (purple color), was able to suppress spontaneous aggregation of 10 μM Aβ 1-42 . D; ThT kinetic graphs showing that aggregation of 10 μM Aβ 1-42 was not altered by the presence of 50 μM lactic acid, 50 μM glycolic acid or a mixture of 50 μM lactic acid + glycolic acid. E; Isothermal titration calorimetry data obtained after base-line subtraction for the titration of 600 μM PLGA with 10 μM Aβ 1-42 at 37 °C. The data were plotted for a single-site binding model and the thermodynamic parameters for the protein and ligand complex interaction are presented in the inset. F; Decreased fluorescence emission of fluorescent PLGA nanoparticles in the presence of Aβ 1-42 monomers revealing the quenching effect during the interaction. G; Epitope mapping using a filter-trap assay revealing that PLGA nanoparticles interact primarily with the hydrophobic domain of Aβ 1-42. H; Epitope mapping of different Aβ 1-42 conformers (i.e., monomer, oligomer and fiber) using filter-trap assay revealing that interaction of 50 μM PLGA with Aβ monomer is somewhat more site-specific than Aβ oligomers and aggregates.

Article Snippet: Dulbecco's modified Eagle's medium (DMEM), neurobasal medium, Hanks' balanced salt solution (HBSS), fetal bovine serum (FBS), Alexa Fluor 488/594 conjugated secondary antibodies, ProLong Gold anti-fade reagent, NuPAGE 4–12% Bis-Tris gels and ELISA kits for the detection of mouse and human Aβ 1–40 and Aβ 1–42 were purchased from R&D systems (Minneapolis, MN, USA).

Techniques: Circular Dichroism, Incubation, Isothermal Titration Calorimetry, Titration, Binding Assay, Fluorescence, TRAP Assay

Journal: Bioactive Materials

Article Title: Significance of native PLGA nanoparticles in the treatment of Alzheimer's disease pathology

doi: 10.1016/j.bioactmat.2022.05.030

Figure Lengend Snippet: Aβ 1-42 binding sites predicted by AutoDock/Chimera and observed after post-docking MD simulations A; Monomer Aβ 1-42 with two PLGA molecules remaining attached and two detached after 30ns of MD simulations. B; Tetramer formed from the solution of eight Aβ peptides and two PLGA molecules attached to it after 30ns long simulations. C; Aβ oligomer with four PLGA molecules bound to it. D; Fibril fragment with four PLGAs attached and two detached after 70ns of MD simulations. PLGA molecules are highlighted in green and residues of Aβ peptides involved in docking are shown in grey. Colored surfaces depict hydrophobicity of Aβ binding cavities (blue - hydrophilicity, red - hydrophobicity). Aβ peptides are shown in ribbon representation. β-strands are indicated with teal color and random coils and turns in grey.

Article Snippet: Dulbecco's modified Eagle's medium (DMEM), neurobasal medium, Hanks' balanced salt solution (HBSS), fetal bovine serum (FBS), Alexa Fluor 488/594 conjugated secondary antibodies, ProLong Gold anti-fade reagent, NuPAGE 4–12% Bis-Tris gels and ELISA kits for the detection of mouse and human Aβ 1–40 and Aβ 1–42 were purchased from R&D systems (Minneapolis, MN, USA).

Techniques: Binding Assay

Journal: Bioactive Materials

Article Title: Significance of native PLGA nanoparticles in the treatment of Alzheimer's disease pathology

doi: 10.1016/j.bioactmat.2022.05.030

Figure Lengend Snippet: Disassembly of aggregated Aβ 1-42 fibers by PLGA A & B; PLGA dose-dependently (2.5–50 μM) triggered disassembly of aggregated Aβ 1-42 as revealed by the ThT fluorescence assay (A) and ThT-stained fluorescence images (B). C; ThT stained images of matured Aβ 1-42 fibers before (i) and after (ii) treatment with 25 μM PLGA revealing the untwining effect following disassembly. D; Fluorescence imaging of Congo red-stained Aβ 1-42 fibers in the presence of green fluorescent PLGA showing a direct interaction between Aβ 1-42 fibers and PLGA (i-iv). E; TEM images of matured Aβ 1-42 fibers in the absence (i) and presence of 50 μM PLGA (ii-iv) depicting the direct interaction of the nanoparticles with Aβ fibers. F–I; DLS analysis revealing the hydrodynamic radius of 10 μM Aβ 1-42 aggregates in the absence (F) and presence of 2.5 μM (G), 25 μM (H) and 50 μM (I) PLGA. Note the shift in the hydrodynamic radius of Aβ 1-42 fibers towards the lower ordered species and possible release of monomers in the presence of PLGA nanoparticles. J; Native PAGE analysis of 10 μM Aβ 1-42 aggregates in the absence and presence of different concentrations (2.5–50 μM) of PLGA. Note the loss of higher-ordered Aβ 1-42 entities following incubation with PLGA. K; Fluorescence quenching effect of labelled PLGA in the presence of increasing concentrations of matured Aβ 1-42 fibers displaying a decrease in fluorescence intensity, suggesting an interaction between PLGA and Aβ 1-42 fibers.

Article Snippet: Dulbecco's modified Eagle's medium (DMEM), neurobasal medium, Hanks' balanced salt solution (HBSS), fetal bovine serum (FBS), Alexa Fluor 488/594 conjugated secondary antibodies, ProLong Gold anti-fade reagent, NuPAGE 4–12% Bis-Tris gels and ELISA kits for the detection of mouse and human Aβ 1–40 and Aβ 1–42 were purchased from R&D systems (Minneapolis, MN, USA).

Techniques: Fluorescence, Staining, Imaging, Clear Native PAGE, Incubation

Journal: Bioactive Materials

Article Title: Significance of native PLGA nanoparticles in the treatment of Alzheimer's disease pathology

doi: 10.1016/j.bioactmat.2022.05.030

Figure Lengend Snippet: Attenuation of Aβ 1-42 aggregation by PLGA protects cultured neurons A & B; Histogram depicting dose-dependent decrease in the viability of mouse cortical cultured neurons following 24h exposure with oligomeric human Aβ 1-42 compared to neurons treated with 10 μM Aβ 42-1 (Cont) as revealed by MTT (A) and LDH (B) assays. C–F; Histograms showing protection of mouse cortical cultured neurons after 24h (C, D) and 100h (E, F) treatment with 10 μM Aβ 1-42 exposed to 25 and 50 μM PLGA as detected with MTT (C, E) and LDH (D, F) assays. G-N; Immunoblots and corresponding histograms showing that protective effects following attenuation of spontaneous Aβ aggregation by PLGA are mediated by decreasing Phospho-Tyr GSK-3β (G, K), Phospho-ERK1/2 (H, L), Phospho-tau (I, M) and cleaved-caspase-3 (J, N) levels. O; LysoSensor labelling of mouse cortical cultured neurons in the absence of Aβ 1-42 (i), after treatment with 10 μM Aβ 1-42 following 24h aggregation (ii), after treatment with 25 μM PLGA (iii) and after treatment with 10 μM Aβ 1-42 incubated with 25 μM PLGA for 24h (iv). Note that attenuation of Aβ aggregation by PLGA partially reversed the basic pH environment observed in Aβ 1-42 -treated neurons. P; Confocal microscopy showing distribution of immunoreactive cathepsin D in the absence of Aβ 1-42 (i), after treatment with 10 μM Aβ 1-42 following 24h aggregation (ii), after treatment with 25 μM PLGA (iii) and after treatment with 10 μM Aβ 1-42 incubated with 25 μM PLGA for 24h (iv). Note that PLGA partially reversed cytosolic cathepsin D labelling in Aβ-treated neurons. All results, which are presented as means ± SEM, were obtained from three to five separate experiments. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Dulbecco's modified Eagle's medium (DMEM), neurobasal medium, Hanks' balanced salt solution (HBSS), fetal bovine serum (FBS), Alexa Fluor 488/594 conjugated secondary antibodies, ProLong Gold anti-fade reagent, NuPAGE 4–12% Bis-Tris gels and ELISA kits for the detection of mouse and human Aβ 1–40 and Aβ 1–42 were purchased from R&D systems (Minneapolis, MN, USA).

Techniques: Cell Culture, Western Blot, Incubation, Confocal Microscopy

Journal: Bioactive Materials

Article Title: Significance of native PLGA nanoparticles in the treatment of Alzheimer's disease pathology

doi: 10.1016/j.bioactmat.2022.05.030

Figure Lengend Snippet: PLGA attenuates AD-related cognitive deficit and pathology in 5xFAD mice A; Photomicrographs depicting OC antibody labelled Aβ deposits in the cortex (i-iv) and cerebellum (v-viii) of PLGA- and CSF-treated WT and 5xFAD mouse brain tissues. As expected, Aβ deposits are evident in the cortex (iii) but not in the cerebellum (vii) of 5xFAD mouse brains. WT mice did not display Aβ deposits in any brain regions (i, v). B-D; Histograms showing the quantification of OC antibody labelled Aβ plaques number (B), size (C) and areas occupied (D) in the cortical (i.e., frontal, parietal and entorhinal) regions of PLGA- and CSF-treated 5xFAD mice. Note the decreased number/size and areas occupied by Aβ-containing neuritic plaques in PLGA-treated 5xFAD mice compared to CSF- treated 5xFAD mice. E-I; Immunoblots (E, F) and their quantification (G–I) showing the levels of APP holoprotein (Y188) and APP-CTFs (i.e., αCTF and βCTF) in the parietal cortex (E) and cerebellum (F) of PLGA- and CSF-treated 5xFAD mouse brain tissues. Note the decreased levels of APP and its cleaved products in the cortex of PLGA-treated 5xFAD mice compared to CSF-treated 5xFAD mice (G–I). The quantifications of the respective blots were represented as a percentage of the corresponding control group (G–I). J & K; Histograms depicting decreased cortical, but not cerebellar, levels of Aβ 1-40 (J) and Aβ 1-42 (K) in PLGA-treated 5xFAD mice compared to CSF-treated 5xFAD mice. L & M; Histograms showing the reversal of object recognition memory following PLGA treatment in 5xFAD mice. The data represent the total number of visits to novel object (L) and the discrimination index (M) from familiar to the novel object of PLGA- and CSF-treated animals. All data expressed as mean ± SEM were obtained from three replicates. CSF, artificial cerebrospinal fluid; WT, wildtype. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Dulbecco's modified Eagle's medium (DMEM), neurobasal medium, Hanks' balanced salt solution (HBSS), fetal bovine serum (FBS), Alexa Fluor 488/594 conjugated secondary antibodies, ProLong Gold anti-fade reagent, NuPAGE 4–12% Bis-Tris gels and ELISA kits for the detection of mouse and human Aβ 1–40 and Aβ 1–42 were purchased from R&D systems (Minneapolis, MN, USA).

Techniques: Western Blot

Journal: Bioactive Materials

Article Title: Significance of native PLGA nanoparticles in the treatment of Alzheimer's disease pathology

doi: 10.1016/j.bioactmat.2022.05.030

Figure Lengend Snippet: PLGA protects iPSC-derived AD neurons against Aβ-induced toxicity A; Representative immunofluorescence images of iPSC-derived neurons from a healthy control (i, ii, v, vi) and a sporadic AD patient (iii, iv, vii, viii) labelled with different neuronal markers. The upper panel shows immunofluorescence labelling with βIII Tubulin (neuronal microtubule marker) (i and iii), βIII Tubulin + SMI312 (pan axonal marker) + DAPI (nuclear marker) (ii and iv), whereas the lower panel reveals immunofluorescence labelling with NeuN (neuronal nuclear marker)(v and vii), NeuN + MAP2 (dendritic marker) + DAPI (vi and viii) in iPSC-derived control and sporadic AD neurons. B & C; Histograms showing the cellular (B) and secretory (C) levels of human Aβ 1-40 in iPSC-derived primary neurons from two control and two sporadic AD-patients. Note the significant increase in cellular Aβ 1-40 levels in both iPSC-AD neurons, while the secretory Aβ 1-40 level was found to be increased only in one case of iPSC-AD neurons compared to controls. D & E; Histograms showing protection of cultured neurons derived from control and sporadic AD following 24h exposure to 5 μM Aβ 1-42 with or without 25 and 50 μM PLGA as detected with MTT (D) and cell toxicity (E) assays. F & G; Histograms showing protection of cultured neurons derived from control and sporadic AD following 100h exposure to 5 μM Aβ 1-42 in the presence or absence of 25 and 50 μM PLGA as detected with MTT (F) and cell toxicity (G) assays. H–O; Immunoblots and corresponding histograms depicting that protective effect of PLGA in one control and sporadic AD neurons against 5 μM Aβ 1-42 following 24h exposure is mediated by decreasing the levels of Phospho-Tyr GSK-3β (H, L), Phospho-ERK1/2 (I, M), Phospho-tau (J, N) and cleaved-caspase-3 (K, O). All data expressed as mean ± SEM were obtained from three-four replicates. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Dulbecco's modified Eagle's medium (DMEM), neurobasal medium, Hanks' balanced salt solution (HBSS), fetal bovine serum (FBS), Alexa Fluor 488/594 conjugated secondary antibodies, ProLong Gold anti-fade reagent, NuPAGE 4–12% Bis-Tris gels and ELISA kits for the detection of mouse and human Aβ 1–40 and Aβ 1–42 were purchased from R&D systems (Minneapolis, MN, USA).

Techniques: Derivative Assay, Immunofluorescence, Marker, Cell Culture, Western Blot