antibody ps129-α-syn (d1r1r)  (Cell Signaling Technology Inc)

Journal: NPJ Parkinson's Disease

Article Title: IC100 blocks inflammasome activation induced by α-synuclein aggregates and ASC specks

doi: 10.1038/s41531-025-00963-8

Figure Lengend Snippet: ASC expression and accumulation vary in different morphologies of Lewy body formation. PD sections presented with different forms of Lewy body pathologies; irregular-shaped ( A ), Lewy bodies ( B ), pale bodies ( C ; black asterisk) and free neuromelanin ( C ; black arrowhead) were stained with human anti-ASC antibody (IC100, black asterisks) in the SN of PD donors and controls ( D and E ). A significant reduction of SN granular was seen in PD donors compared to controls ( F ). In ( G ), IC100 immunostaining is present as granular/widespread in dopaminergic neurons (blue arrow) or within the intracellular Lewy body formation (white asterisk). Mouse anti-α-synuclein (Clone 42) is present in Lewy body fragments ( Gi–Giii ); white (red) asterisk marks where the Lewy body formed in neuron seen in image ( G ) in neurons and in ameboid-shaped microglia ( Gii , green; yellow arrow). H IC100 (red) distribution in control dopaminergic neurons in the SN. α-Synuclein staining in white matter tracts ( Hi ; white) but not in microglia ( Hii ; green) or neurons that show granular IC100 immunostaining ( Hiii ; green). NLRP1 is present in Lewy body clusters ( I ) in donors with PD pathology, but not in controls ( K ). IC100 colocalized with NLRP1 ( J ; Blue arrow and Ji ) in neurons that were positive for pSer129 α-synuclein ( Jii ). L , Li , and Lii correspond to controls that present few neurons with cytoplasmic IC100. mm = millimeter; ** p < 0.01; α-Syn = alpha synuclein; Adapter protein apoptosis-associated speck-like protein containing a caspase recruitment domain (ASC); Substantia nigra (SN). Scale bars = 10 μm ( A – E , Gi – Giii , Hi – Hiii , I , Ji , Jii , K , Li , and Lii ) and 20 μm ( G , H , J , and L ).

Article Snippet: Sections were then blocked in 5% goat serum (Vector Laboratories, Burlingame, Calif., USA) for 20 min before being incubated overnight at 4 °C in a solution of rabbit anti-Phospho-α-Synuclein (Ser129) (D1R1R) antibody (D1R1R; 0.3 μg/mL; Cell Signaling Technologies Danvers, MA, USA), rabbit anti-NLRP3 (3.0 μg/mL; MilliporeSigma, St. Louis, MO, USA), mouse anti-NLRP1 (1.0 μg/mL; Enzo Life Sciences, Farmingdale, NY., USA), or human anti-ASC (IC100, 2 μg/mL; as described in refs. , ) in PBS.

Techniques: Expressing, Staining, Immunostaining, Control

Journal: eNeuro

Article Title: Alpha-Synuclein Phosphomimetic Y39E and S129D Knock-In Mice Show Cytosolic Alpha-Synuclein Localization without Developing Neurodegeneration or Motor Deficits

doi: 10.1523/ENEURO.0357-24.2025

Figure Lengend Snippet: Materials used for this study

Article Snippet: p-S129-α-Syn (clone D1R1R) , Cell Signaling Technology , RRID: AB_279886.

Techniques: Protease Inhibitor, Western Blot, Immunodetection, Plasmid Preparation, CRISPR, Sequencing, Genomic Sequencing

Journal: medRxiv

Article Title: Progression GWAS followed by functional characterization implicates E3 ubiquitin ligase TRIM2 as a potential genetic modifier of Parkinson’s disease progression

doi: 10.1101/2025.02.21.25322301

Figure Lengend Snippet: a. 15 pairwise differential analyses of time-course transcriptomes in neurons that underwent 21 days of a-syn aggregation identify >3,000 PFF-induced DEGs. 3 key comparisons are shown here. PBS is baseline timepoint before PFF treatment. D: days passed after PFF-treatment. NS: Not significant. FC: fold-change. b. 3-way comparison of AMP-PD and PPMI genes with suggestive association (P < 1E-5) and PFF-induced DEGs identify 137 candidate genes for PD progression supported by 2+ datasets. c. Trim2 shows consistent decrease in expression over the course of a-syn aggregation. In 10/15 comparisons, the differential expression was significant. Med13l shows a trend of increased expression after pS129+ aggregate observation at D4, with a statistically significant increase at the timepoints (D21, D14, and D7) vs. PBS and D1 time points and at D3 vs. PBS. Ppp2r3a has decreased expression with increasing length of PFF treatment, with significant changes at D3, D7, D14, and D21 vs. PBS. Glb1l2 has decreased expression after 3 days of PFF treatment, then increased again with increasing length of PFF treatment, with significant changes at D21 vs. all other timepoints, D3 vs. D1 and PBS, and D14 vs. D3.

Article Snippet: Primary antibodies used for ICC include: pS129 α-syn [D1R1R] rabbit mAb (Cell Signaling Technology, Cat# 23706, 1:3000), MAP2 chicken antibody (abcam, Cat# ab92434, 1:5000), anti-NeuN mouse mAb, [clone A60] (Millipore, Cat# MAB377, 1:2000), and Neurofilament-L [DA2] mouse mAb (Cell Signaling Technology, Cat# 2835, 1:2000).

Techniques: Comparison, Expressing

Journal: medRxiv

Article Title: Progression GWAS followed by functional characterization implicates E3 ubiquitin ligase TRIM2 as a potential genetic modifier of Parkinson’s disease progression

doi: 10.1101/2025.02.21.25322301

Figure Lengend Snippet: CD1 mouse primary neurons were cultured for 21 days in vitro (DIV) with or without Accell siRNA knockdown of murine TRIM2 and treated with 1 µg/ml human α-syn preformed fibrils (PFFs) for the final 7 or 14 days. a. Representative images of DIV 21 neurons treated with a-syn PFFs for 14d and stained for DAPI (blue), neurofilament-L (NF-L, green), and pS129 α-syn (red). Arrows denote inclusions that are co-positive for NF-L and pS129 α-syn. Scale bar = 10µm. b-c. Quantification of the percentage of pS129 a-syn aggregates that are co-positive for NF-L (b) and total NF-L intensity per MAP2+ area (c) in a. *p=0.0141 by unpaired Student’s t-test; n=6wells/group. d. Representative images of neuronal cell body pS129 α-syn aggregates (red, arrows) and NF-L (green, bottom panel) in neurons treated with 1 µg/ml PFFs for 14d. Note the accumulation of NF-L in neurons treated with TRIM2 siRNAs. Scale bar = 5µm. e . Representative images of DAPI (blue), MAP2 (orange), and pS129 α-syn (red) immunostaining in DIV 21 CD1 neurons treated with TRIM2 or non-targeting control (CTRL) siRNAs and 1 µg/ml PFFs for 14d. Scale bar = 20µm. f-g. Quantification of total pS129 α-syn pathology per MAP2+ area in (e). after 7d or 14d treatment with PFFs for CD1 neurons treated with TRIM2 siRNAs (f) or human TRIM2 lentivirus (hTRIM2 LV, g). ****p<0.0001 *p=0.0313 by 2-Way ANOVA with Sidak’s multiple comparisons test; n=9 wells/group. Figures a-g include data from a single representative experiment repeated at least 3 times in independent cultures of primary mouse neurons. Error bars represent mean ± SEM.

Article Snippet: Primary antibodies used for ICC include: pS129 α-syn [D1R1R] rabbit mAb (Cell Signaling Technology, Cat# 23706, 1:3000), MAP2 chicken antibody (abcam, Cat# ab92434, 1:5000), anti-NeuN mouse mAb, [clone A60] (Millipore, Cat# MAB377, 1:2000), and Neurofilament-L [DA2] mouse mAb (Cell Signaling Technology, Cat# 2835, 1:2000).

Techniques: Cell Culture, In Vitro, Knockdown, Staining, Immunostaining, Control

Journal: medRxiv

Article Title: Progression GWAS followed by functional characterization implicates E3 ubiquitin ligase TRIM2 as a potential genetic modifier of Parkinson’s disease progression

doi: 10.1101/2025.02.21.25322301

Figure Lengend Snippet: Neurofilament-L (NF-L) and α-syn both accumulate in PD, with a significant portion of α-syn and NF-L aggregates co-localizing both in neurites and in the cell body. TRIM2 ubiquitinates NF-L protein, thereby targeting it for proteasomal degradation and promoting normal protein homeostasis (left panel). Loss of TRIM2 function results in aberrant accumulation of NF-L which may serve to further promote α-syn aggregation, inhibit α-syn clearance by other mechanisms, or both (right panel). Therefore, activation of TRIM2 activity or upregulation of TRIM2 protein level may slow the progression of PD by both reducing α-syn pathology and promoting normal cellular homeostasis in neurons.

Article Snippet: Primary antibodies used for ICC include: pS129 α-syn [D1R1R] rabbit mAb (Cell Signaling Technology, Cat# 23706, 1:3000), MAP2 chicken antibody (abcam, Cat# ab92434, 1:5000), anti-NeuN mouse mAb, [clone A60] (Millipore, Cat# MAB377, 1:2000), and Neurofilament-L [DA2] mouse mAb (Cell Signaling Technology, Cat# 2835, 1:2000).

Techniques: Activation Assay, Activity Assay