Journal: Acta Neuropathologica Communications

Article Title: Altered amyloid-β structure markedly reduces gliosis in the brain of mice harboring the Uppsala APP deletion

doi: 10.1186/s40478-024-01734-x

Figure Lengend Snippet: Aβ immunostaining and ELISA-based measurements of Aβ aggregates in brain tissue from tg-UppSwe, tg-ArcSwe and tg-Swe mice. A Immunostaining of tg-UppSwe, tgArcSwe and tg-Swe brain tissue sections (hippocampus, 40× magnification) with 3D6 (yellow, upper panel) or mAb158 (yellow, lower panel) in comparison with double staining using Aβ40 and Aβ42-specific antibodies (green). Scale bar: 200 µm. B ELISA quantification of total Aβ1-40 and 1-42 levels in FA brain extract. C 3D6-3D6 ELISA quantification of total Aβ aggregates in TBS 100K , TBS 16K and TBS-T brain extracts. D mAb158 positive fraction of the total Aβ aggregates detected by 3D6-3D6 ELISA in TBS 100K , TBS 16K and TBS-T brain extracts. E Distribution of [ 125 I]RmAb3D6-scFv8D3 and [ 125 I]RmAb158-scFv8D3, three days after administration of these bispecific antibodies at 32 nmol/kg body weight (therapeutic dose) to 18-month-old tg-UppSwe mice, expressed as a brain-to-blood radioactivity ratio in whole brain tissue (Brain) and in TBS 16K , TBS-T and FA extracts. F Soluble Aβ aggregates in TBS 100K and TBS 16 K brain extracts from 18 months old tg-UppSwe mice three days after administration of a therapeutic dose (32 nmol/kg) of [ 125 I]RmAb3D6-scFv8D3 or [ 125 I]RmAb158-scFv8D3, in comparison with PBS. Non-significant ( ns ), * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: The TBS 16K and TBS-T brain extracts were diluted in ELISA incubation buffer (PBS, 0.1% BSA, 0.05% Tween) and incubated over night at 4 °C, followed by detection with 0.25 μg/ml biotinylated anti-TREM2 BAF1729 (R&D), HRP-conjugated streptavidin and K Blue Aqueous TMB substrate and read at 450 nm with a spectrophotometer.

Techniques: Immunostaining, Enzyme-linked Immunosorbent Assay, Comparison, Double Staining, Radioactivity

Journal: Acta Neuropathologica Communications

Article Title: Altered amyloid-β structure markedly reduces gliosis in the brain of mice harboring the Uppsala APP deletion

doi: 10.1186/s40478-024-01734-x

Figure Lengend Snippet: Astroglial response to Aβ. A Aβ40 + 42 and GFAP immunostaining of brain tissue from 18-month-old tg-UppSwe, tg-ArcSwe and tg-Swe mice. Co-localization of Aβ and GFAP was lacking in tg-UppSwe mice but was abundant in tg-ArcSwe and tg-Swe mice. Scale bar 100 µm. ELISA quantification of GFAP in TBS ( B ) and TBS-T ( C ) brain extracts from 18-months-old wt, tg-UppSwe, tg-ArcSwe and tg-Swe mice. Not significant ( ns ), ** P < 0.01, *** P < 0.001. D Schematic description of the procedure to establish primary astrocyte monocultures originating from the cerebral cortices of embryonic mouse brain. Primary astrocyte cultures were exposed to sonicated, Cy3-labeled fibrils of synthetic Aβ Upp , Aβ Arc or Aβ wt (all Aβ1-42). Cells were stained for GFAP, LAMP-1 and cell nuclei (Dapi). While Aβ Upp clustered on the surface of cells, Aβ Arc and Aβ wt appeared to be phagocytosed to a larger extent. Scale bar: 20 µm

Article Snippet: The TBS 16K and TBS-T brain extracts were diluted in ELISA incubation buffer (PBS, 0.1% BSA, 0.05% Tween) and incubated over night at 4 °C, followed by detection with 0.25 μg/ml biotinylated anti-TREM2 BAF1729 (R&D), HRP-conjugated streptavidin and K Blue Aqueous TMB substrate and read at 450 nm with a spectrophotometer.

Techniques: Immunostaining, Enzyme-linked Immunosorbent Assay, Sonication, Labeling, Staining

Journal: Acta Neuropathologica Communications

Article Title: Altered amyloid-β structure markedly reduces gliosis in the brain of mice harboring the Uppsala APP deletion

doi: 10.1186/s40478-024-01734-x

Figure Lengend Snippet: Microglial response to Aβ. A Aβ, TREM2 and Iba-1 immunostaining of brain tissue from 18-months-old tg-UppSwe, tg-ArcSwe and tg-Swe mice. Scale bar: 100 µm. ELISA quantification of soluble TREM2 in TBS 16K ( B ) and TBS-T ( C ) brain extracts from 18-months-old wt, tg-UppSwe, tg-ArcSwe and tg-Swe mice. Not significant ( ns ), * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: The TBS 16K and TBS-T brain extracts were diluted in ELISA incubation buffer (PBS, 0.1% BSA, 0.05% Tween) and incubated over night at 4 °C, followed by detection with 0.25 μg/ml biotinylated anti-TREM2 BAF1729 (R&D), HRP-conjugated streptavidin and K Blue Aqueous TMB substrate and read at 450 nm with a spectrophotometer.

Techniques: Immunostaining, Enzyme-linked Immunosorbent Assay

Journal: Pharmaceutical Research

Article Title: Transferrin Receptor Binding BBB-Shuttle Facilitates Brain Delivery of Anti-Aβ-Affibodies

doi: 10.1007/s11095-022-03282-2

Figure Lengend Snippet: Indirect ELISA of the four affibodies and bispecific Aβ-antibody fusion protein di-scFv3D6-8D3. Plates were coated with ( a ) 250 nM Aβ protofibrils ( b ) 2 μg/ml mTfR1.

Article Snippet: Affibody and secondary antibody sample dilutions were made in ELISA incubation buffer (PBS, 0.1% BSA, 0.05% Tween-20).

Techniques: Indirect ELISA

Journal: Pharmaceutical Research

Article Title: Transferrin Receptor Binding BBB-Shuttle Facilitates Brain Delivery of Anti-Aβ-Affibodies

doi: 10.1007/s11095-022-03282-2

Figure Lengend Snippet: Affinities (K D ) in nM for Aβ-Protofibrils and mTfR1 Determined by Indirect ELISA for the Affibodies and di-scFv3D6-8D3. Mean Fold Difference in K D Between Unlabeled and 125 I-Labeled Affibody and di-scFv3D6-8D3, Towards Aβ-protofibrils and mTfR1 Determined by Indirect ELISA (Values Expressed as Mean ± SD)

Article Snippet: Affibody and secondary antibody sample dilutions were made in ELISA incubation buffer (PBS, 0.1% BSA, 0.05% Tween-20).

Techniques: Indirect ELISA, Radioactivity