biotinylated goat antimouse igm  (Vector Laboratories)

Journal: Alzheimer's Research & Therapy

Article Title: Anti-β-sheet conformation monoclonal antibody reduces tau and Aβ oligomer pathology in an Alzheimer’s disease model

doi: 10.1186/s13195-018-0337-3

Figure Lengend Snippet: Protocol of infusion of GW-23B7 or vehicle control into triple-transgenic (3 × Tg) mice and the ensuing behavioral and kinetic tests. a Protocol of the intraperitoneal infusion of GW-23B7 or vehicle alone into 3 × Tg Alzheimer’s disease mice, behavioral tests, and killing of the tested animals. b Comparative kinetics of pentameric immunoglobulin M (IgM) distribution inside brains pooled from 18-month-old 3 × Tg animals infused with either GW-23B7 or vehicle alone as per the bottom part of protocol in ( a ). Western blots show soluble supernatants from 20% brain homogenates that were stained before and after reduction with reversible Fast Green to assess comparable protein loading ( top panel ) and detected by antimouse μ heavy chain or antimouse kappa light chain ( middle and bottom panels , respectively). The relative concentrations determined by densitometry of the bands are plotted on the graph on the right. c Radial arm maze behavioral test showing significant differences ( p < 0.0001 by two-way analysis of variance [ANOVA]) between Tg animals infused with GW-23B7 ( n = 10) and both Tg vehicle-treated 19-month-old ( n = 10) Tg mice at the baseline age of 16–17 months ( n = 9). Dunnett’s multiple comparisons test showed no significant difference between the Tg mice at the baseline age of 16–17 months and the Tg vehicle-treated 19-month-old mice. There was also a significant time learning effect ( p < 0.0001 by two-way ANOVA). DTT Dithiothreitol

Article Snippet: Slides were washed three times with PBS and incubated with biotinylated antimouse IgM (μ-specific chain) antibody (Vector Laboratories, Burlingame, CA, USA) diluted 1:1000 in PBS for 1 h, followed by 1-h incubation in VECTASTAIN® AB solution (Vector Laboratories).

Techniques: Transgenic Assay, Western Blot, Staining

Journal: Alzheimer's Research & Therapy

Article Title: Anti-β-sheet conformation monoclonal antibody reduces tau and Aβ oligomer pathology in an Alzheimer’s disease model

doi: 10.1186/s13195-018-0337-3

Figure Lengend Snippet: Subcloned and purified anti-β-sheet conformational monoclonal antibody (aβComAb) GW-23B7-specific reactivity to human paired helical filaments (PHFs) and oligomeric Aβ peptides. a Enzyme-linked immunosorbent assay (ELISA) data showing the reactivity from cell supernatant of subcloned GW-23B7 immunoglobulin M (IgM) and an irrelevant clone from the same fusion to PHFs and oligomers differential on Aβ 1–40 and Aβ 1–42 . The right panel shows the even coating of the selected peptides on the plate and the lack of unspecific reactivity to secondary antimouse IgM. b Western blots showing the pentameric integrity of the purified aβComAb GW-23B7. Lane 1 = unreduced sample, lane 2 = reduced with 0.1 M dithiothreitol (DTT). Left panel : Fast Green reversible protein stain. Middle panel : Antimouse IgM (μ-specific) light chain. Right panel : Antimouse kappa light chain. IgMp Pentameric immunoglobulin M, Hμr μ heavy chain reduced, Kr Kappa light chain reduced. c ELISA results showing the reactivity of purified aβComAb GW-23B7 diluted 1:1000 to Aβ 1–40 , Aβ 1–42 , and human PHFs. d Surface plasmon resonance showing the binding affinity of the purified aβComAb GW-23B7 to the oligomeric species of Aβ 1–42 and the lack of binding affinity to the monomeric forms. The dissociation constant (K D ) (14 nM) was determined from the raw data on the left

Article Snippet: Slides were washed three times with PBS and incubated with biotinylated antimouse IgM (μ-specific chain) antibody (Vector Laboratories, Burlingame, CA, USA) diluted 1:1000 in PBS for 1 h, followed by 1-h incubation in VECTASTAIN® AB solution (Vector Laboratories).

Techniques: Purification, Enzyme-linked Immunosorbent Assay, Western Blot, Staining, SPR Assay, Binding Assay