Journal: bioRxiv

Article Title: Plaque-associated endogenous IgG and its impact on immunohistochemical detection of mouse monoclonal IgG antibodies in mouse models of Alzheimer’s disease

doi: 10.1101/2024.04.25.591057

Figure Lengend Snippet: ( a , b ) Applications of an AF 488 conjugated Dk anti-Ms IgG (H+L) (yellow) on App NL-G-F/NL-G-F mouse brain sections pretreated with Fab fragments of a Dk anti-Ms IgG (H+L) Ab (n = 3 mice for each condition). 10 ( a ) and 1.0 ( b ) µg/mL of the secondary Ab are applied. Aβ plaques are labeled with FSB (cyan). The top and bottom panels show images of the secondary Ab, and merged images of the secondary Ab and FSB. Concentrations of the secondary Ab and Fab fragments are indicated on the top. The percentages of Aβ plaques decorated with the secondary Ab are shown in the bottom left corner of the bottom panels. Double arrowheads and arrowheads indicate off-target binding of the secondary Ab and Aβ plaques, respectively. Scale bar, 20 µm.

Article Snippet: The number of Aβ plaques decorated with an AF 488 Dk anti-Ms IgG (H+L) Ab (A21202, Thermo Fisher Scientific) among total Aβ plaques was counted, and the proportion of these Aβ plaques in the total number of Aβ plaques was calculated.

Techniques: Labeling, Binding Assay

Journal: bioRxiv

Article Title: Plaque-associated endogenous IgG and its impact on immunohistochemical detection of mouse monoclonal IgG antibodies in mouse models of Alzheimer’s disease

doi: 10.1101/2024.04.25.591057

Figure Lengend Snippet: ( a - c ) Applications of AF 488 conjugated nAbs against Ms IgG1 ( a ), 2b ( b ), and IgGκ light chain ( c ) (yellow) on frozen brain sections of App NL-G-F/NL-G-F mice (n = 3 mice for each condition). Aβ plaques are labeled with FSB (cyan). Images of secondary nAbs, and merged images of the secondary nAbs and FSB are shown in the left and right panels, respectively. Double arrowheads and arrowheads indicate off-target binding of the secondary nAb and Aβ plaques, respectively. Scale bar, 20 µm.

Article Snippet: The number of Aβ plaques decorated with an AF 488 Dk anti-Ms IgG (H+L) Ab (A21202, Thermo Fisher Scientific) among total Aβ plaques was counted, and the proportion of these Aβ plaques in the total number of Aβ plaques was calculated.

Techniques: Labeling, Binding Assay

Journal: bioRxiv

Article Title: Plaque-associated endogenous IgG and its impact on immunohistochemical detection of mouse monoclonal IgG antibodies in mouse models of Alzheimer’s disease

doi: 10.1101/2024.04.25.591057

Figure Lengend Snippet: ( a ) Indirect IF of 4G8 Ab (yellow) using secondary Abs against Ms IgG (H+L) (left) and IgG2b (right) on frozen brain sections of App NL-G-F/NL-G-F mice (n = 3 mice for each condition). Aβ plaques are labeled with FSB and indicated by arrowheads (cyan). ( b ) Triple labeling of 4G8 Ab (yellow), an Iba1 Ab (magenta), and FSB (cyan) on frozen brain sections of App NL-G-F/NL-G-F mice (n = 3 mice for each condition). AF 488 conjugated anti-Ms IgG (H+L) (left) and Ms IgG2b Abs (right) are used as secondary Abs. Triple merged images of 4G8 Ab, the Iba1 Ab, and FSB are shown in the bottommost panel. Double arrowheads indicate off-target binding to PA-IgG. ( c ) Triple labeling of AT8 Ab (yellow), the Iba1 Ab (magenta), and FSB (cyan) on frozen brain sections of App NL-G-F/NL-G-F mice (n = 3 mice for each condition). AT8 Ab is indirectly detected with secondary Abs against IgG (H+L) (left) and IgG1 (right). The bottommost panel show triple merged images of AT8 Ab, the Iba1 Ab, and FSB. Double arrowheads and arrows show off-target binding to PA-IgG, and immunosignals in dystrophic neurites, respectively. Scale bars, 50 µm in ( a ) and 20 µm in ( c ).

Article Snippet: The number of Aβ plaques decorated with an AF 488 Dk anti-Ms IgG (H+L) Ab (A21202, Thermo Fisher Scientific) among total Aβ plaques was counted, and the proportion of these Aβ plaques in the total number of Aβ plaques was calculated.

Techniques: Labeling, Binding Assay

Journal: medRxiv

Article Title: Autoantibody screening in Guillain-Barré Syndrome

doi: 10.1101/2021.05.10.21256964

Figure Lengend Snippet: DRG neurons ( A-H ) or neuroblastoma neurons ( I-P ) were stained with control or patient’s sera in green. Signal intensity of IgG ( A-D, I-L ) or IgM ( E-H, M-P ) reactivity was scored in a 0-3 scale (0=negative, 1=mild positive, 2=moderate positive, 3=strong positive). Neuroblastoma neurons were counterstained in red with anti-panNeurofascin mAb.

Article Snippet: Goat antihuman IgG (or IgM) AF488 (Molecular probes, Oregon, USA) were used as secondary antibodies at 1/1000 concentration.

Techniques: Staining

Journal: medRxiv

Article Title: Autoantibody screening in Guillain-Barré Syndrome

doi: 10.1101/2021.05.10.21256964

Figure Lengend Snippet: DRG neurons (A-B) stained with a GBS patient’s serum reacting moderately (score 2) in IgG (A) , and a GBS patient’s serum reacting strongly (score 3) in IgM (B) . Human neuroblastoma-derived neurons (C-D) stained in red with anti-panNeurofascin mAb, and in green with a GBS patient’s serum reacting moderately (score 2) in IgG (C) and a GBS patient’s serum reacting strongly (score 3) in IgM (D) .

Article Snippet: Goat antihuman IgG (or IgM) AF488 (Molecular probes, Oregon, USA) were used as secondary antibodies at 1/1000 concentration.

Techniques: Staining, Derivative Assay