Journal: medRxiv

Article Title: Alpha-synuclein misfolding as a fluid biomarker for Parkinson’s disease and synucleinopathies measured with the iRS platform

doi: 10.1101/2024.09.02.24312694

Figure Lengend Snippet: iRS-surface characterization by synthetic αSyn antigens. Panel A : The secondary structure sensitive Amide-I band absorbance of capture-antibody-bound αSyn-monomers (green, Stressmarq Bioscience Inc SPR-321), αSyn-oligomers (orange, Stressmarq Bioscience Inc SPR-466), and αSyn-PPFs (red, Stressmarq Bioscience Inc SPR-322) in PBS at high concentration of 500 ng/ml and scaled (x1.5-4) for comparison. Their structural differences are indicated by the significant wavenumber shift (cm -1 ) ranging from 1652 cm -1 for α-helical/random-coil (green) over 1647 cm -1 (orange) to β-sheet dominated structures absorbing at 1624 cm -1 (red). Panel B : The inertness measures on the blocking solution (BS) layer at high concentrations (500-5000 ng/ml) without the capture antibody. No signal is observed without the antibody on the blocking layer demonstrating sufficient inertness for pg-ng/ml concentrations of αSyn in CSF. Abbreviations: αSyn, alpha-synuclein; BS, blocking solution; PFF, pre-formed fibril.

Article Snippet: Briefly, αSyn stocks (Stressmarq, Cat. No. SPR-321), stored at -80 °C, were slowly thawed on ice.

Techniques: Concentration Assay, Comparison, Blocking Assay

Journal: Open Forum Infectious Diseases

Article Title: P-1810. Development of Avian and Human Influenza Analytical Reference Materials for Diagnostics and Surveillance

doi: 10.1093/ofid/ofaf695.1979

Figure Lengend Snippet: pdm09 qPCR data Figure 4: qPCR amplification curves generated with synthetic RNA ATCC® VR-3388SD™ (H1N1 pdm09) and genomic RNA samples derived from two viral cultures ATCC® VR-3441 ™ VR-1988™, which are also both of H1N1 pdm09 type, using the WHO assay for the detection of Influenza type A subtype H1pdm09.

Article Snippet: Figure 3: H9N2 qPCR data Figure 3: qPCR amplification curves generated with ATCC® VR-3440SDTM (subtype H9N2) using (A) a Hassan et al., 2022 assay targeting HA, and the (B) CDC Flu SC2 Multiplex assay targeting M. Figure 4: pdm09 qPCR data Figure 4: qPCR amplification curves generated with synthetic RNA ATCC® VR-3388SDTM (H1N1 pdm09) and genomic RNA samples derived from two viral cultures ATCC® VR-3441 TM VR-1988TM, which are also both of H1N1 pdm09 type, using the WHO assay for the detection of Influenza type A subtype H1pdm09.

Techniques: Amplification, Generated, Derivative Assay

Journal: NPJ Parkinson's Disease

Article Title: Glycation modulates glutamatergic signaling and exacerbates Parkinson’s disease-like phenotypes

doi: 10.1038/s41531-022-00314-x

Figure Lengend Snippet: Wild-type littermate (WT) and Thy1-aSyn transgenic mice received an intracerebroventricular (ICV) injection of MGO or vehicle (PBS) and protein brain extracts from several regions analyzed 5 weeks post injection. Protein extracts were resolved by SDS-PAGE or loaded into membranes in a dot-blot system (see Supplementary Fig. ). Membranes were probed with anti-aSyn, anti-CEL, anti-pS129-aSyn and anti-β-actin for normalization. Representative blots showing two samples from each experimental group are shown for aSyn, and densitometric analysis represented for aSyn and CEL in a , b midbrain, c , d striatum, e , f cerebellum, g , h prefrontal cortex, and i , j hippocampus, respectively. k Representative blot showing four samples from vehicle- and MGO-injected Thy1-aSyn mice are shown for pS129-aSyn and aSyn total signal. Densitometric analysis of the ratio between pS129-aSyn and aSyn is presented. l Representative blot showing aSyn probing in soluble and insoluble fraction from vehicle- and MGO-injected Thy1-aSyn mice. Densitometric analysis of the ratio between the insoluble and the sum of both soluble and insoluble signals is presented. At least n = 5 in all groups, data in all panels are average with error bars representing standard deviation, Ordinary one-way ANOVA, * p < 0.05, ** p < 0.01, **** p < 0.0001; unpaired two-tailed t -test with equal SD, # p < 0.05.

Article Snippet: Membranes were incubated with blocking solution (5% bovine serum albumin) in 1× TBS (20 mM Tris, 136 mM NaCl, pH 7.6) at room temperature for 30 min. Primary antibody incubations were carried out overnight at 4 °C, using given concentrations in blocking solution: CEL (Mouse Anti-N ε -carboxyethyl lysine, in a dilution of 1:1000 in blocking solution, Cosmo-Bio, USA), aSyn (Purified Mouse Anti-α-Synuclein antibody, in a dilution of 1:1000 in blocking solution, BD Biosciences; San Jose, CA, USA), phosphorylated aSyn at residue 129 (pS129, [J18] SMC-600, StressMarq; Victoria, Canada), glyoxalase I (Rabbit polyclonal anti-Glyoxalase I antibody – FL-184, sc67351, Santa Cruz Biotechnology), and β-actin (Mouse Monoclonal anti-β-actin antibody in a dilution of 1:5000 in blocking solution, Ambion, Thermo Fisher Scientific; Waltham, MA, USA).

Techniques: Transgenic Assay, Injection, SDS Page, Dot Blot, Standard Deviation, Two Tailed Test