rabbit monoclonal anti p α synuclein  (Cell Signaling Technology Inc)

Journal: NPJ Parkinson's Disease

Article Title: LRRK2 R1627P mutation amplifies environmental risk factors induced chronic inflammation and α-synuclein aggregation in the gut of rats

doi: 10.1038/s41531-026-01281-3

Figure Lengend Snippet: a Volcano plot comparing DEGs for RNA-seq in LRRK2 R1627P vs WT, LRRK2 −/− vs WT ( p value < 0.05, fold change >1.2), n = 3. b BP categories in GO enrichment analysis and KEGG pathway of DEGs in LRRK2 R1627P vs WT, n = 3. c BP categories in GO enrichment analysis and KEGG pathway of DEGs in LRRK2 −/− vs WT, n = 3. d Western blot analysis for TLR4, MyD88 and NF-κB protein levels in the small intestine of rats, n = 4. One-way ANOVA was used. e TEM images of rats small intestinal LP. Ec enterocytes, LP lamina propria. Scale bars 5 μm, n = 3. One-way ANOVA was used. f Western blot analysis for CD68 and CD163 protein levels in the small intestine of rats, n = 4. One-way ANOVA was used. g Immunofluorescence staining images of CD68 (green) and CD163 (red) (upper panel), IL-6 (green) and iNOS (red) (down panel) positive cells in the small intestinal LP of rats. Nuclei were stained with DAPI (blue). Scale bar 100 μm, n = 4. One-way ANOVA was used. h Flow cytometry scatter plot of small intestianl LP isolated from rats, n = 3. CD11b + CD68 + CD163 - cells represent M1 population and CD11b + CD68 - CD163 + cells represent M2 population. i Statistical analysis of total macrophages, M1 macrophages and M2 macrophages in the small intestianl LP using flow cytometry. One-way ANOVA was used. j Immunofluorescence staining images of CD68 (green) and p S129 -α-Syn (red) positive cells in the small intestinal LP of rats. Nuclei were stained with DAPI (blue). Scale bar 100 μm, n = 4. k Statistical analysis of immunofluorescence staining. One-way ANOVA was used. Data are presented as mean ± SEM, * p < 0.05, ** p < 0.01.

Article Snippet: Antibody information: LRRK2 (Abcam, ab133474, 1:200), βIII-Tubulin (Servicebio, GB15139, 1:500), CD68 (Santa Cruz Biotechnology, sc-20060, 1:100), CD163 (Servicebio, GB11340-1, 1:200), iNOS (Affinity Biosciences, AF0199, 1:200), IL-6 (Cell Signaling Technology, D5W4V, 1;200), p S129 -α-Syn (Cell Signaling Technology, 23706S, 1:200).

Techniques: RNA Sequencing, Western Blot, Immunofluorescence, Staining, Flow Cytometry, Isolation

Journal: NPJ Parkinson's Disease

Article Title: LRRK2 R1627P mutation amplifies environmental risk factors induced chronic inflammation and α-synuclein aggregation in the gut of rats

doi: 10.1038/s41531-026-01281-3

Figure Lengend Snippet: a HE staining images of the small intestine of LPS-treated rats. The green circles in the lower panels highlight the red blood cells in the LP. Scale bar 100 μm, n = 4. b Images of goblet cells stained with AB-PAS in the small intestine of LPS-treated rats. Scale bar 100 μm, n = 4. c Immunohistochemical staining images of Cleaved caspase 3 in the small intestine of LPS-treated rats. Arrows show apoptotic IECs. Scale bar 100 μm, n = 4. d Western blot analysis for ZO-1, E-cadherin, TLR4, MyD88, NF-κB, CD68 and CD163 protein levels in the small intestine of LPS-treated rats, n = 4. e Immunofluorescence staining images and quantification of CD68/IL-6 (green) and CD163/iNOS/p S129 -α-Syn (red) positive cells in the small intestinal LP of LPS-treated rats. Nuclei were stained with DAPI (blue). Scale bar 100 μm, n = 4. One-way ANOVA was used. Data are presented as mean ± SEM, * p < 0.05, ** p < 0.01.

Article Snippet: Antibody information: LRRK2 (Abcam, ab133474, 1:200), βIII-Tubulin (Servicebio, GB15139, 1:500), CD68 (Santa Cruz Biotechnology, sc-20060, 1:100), CD163 (Servicebio, GB11340-1, 1:200), iNOS (Affinity Biosciences, AF0199, 1:200), IL-6 (Cell Signaling Technology, D5W4V, 1;200), p S129 -α-Syn (Cell Signaling Technology, 23706S, 1:200).

Techniques: Staining, Immunohistochemical staining, Western Blot, Immunofluorescence

Journal: NPJ Parkinson's Disease

Article Title: LRRK2 R1627P mutation amplifies environmental risk factors induced chronic inflammation and α-synuclein aggregation in the gut of rats

doi: 10.1038/s41531-026-01281-3

Figure Lengend Snippet: a , b Western blot analysis for TLR4, MyD88, NF-κB, CD68 and CD163 protein levels in the small intestine of WT, LRRK2 R1627P and LRRK2 −/− rats treated with saline (control) or TLR4 inhibitor (TAK-242), n = 4. One-way ANOVA was used. c Immunofluorescence staining images and quantification of CD68/IL-6 (green) and CD163/iNOS/p S129 -α-Syn (red) positive cells in the small intestinal LP of rats treated with saline (control) or TLR4 inhibitor (TAK-242). Nuclei were stained with DAPI (blue). Scale bar 100 μm, n = 4. Two-way ANOVA was used; for CD68/IL-6/iNOS analysis interaction p < 0.01; for CD163 analysis interaction p > 0.05; for p S129 -α-Syn analysis interaction p < 0.001. d GO enrichment analysis for DEGs in ConRP vs ConWT, TAKRP vs ConRP, TAKRP vs ConWT, n = 3. e Heatmap of DEGs from ConWT, ConRP and TAKRP group, n = 3. Data are presented as mean ± SEM, * p < 0.05, ** p < 0.01.

Article Snippet: Antibody information: LRRK2 (Abcam, ab133474, 1:200), βIII-Tubulin (Servicebio, GB15139, 1:500), CD68 (Santa Cruz Biotechnology, sc-20060, 1:100), CD163 (Servicebio, GB11340-1, 1:200), iNOS (Affinity Biosciences, AF0199, 1:200), IL-6 (Cell Signaling Technology, D5W4V, 1;200), p S129 -α-Syn (Cell Signaling Technology, 23706S, 1:200).

Techniques: Western Blot, Saline, Control, Immunofluorescence, Staining

Journal: NPJ Parkinson's Disease

Article Title: LRRK2 R1627P mutation amplifies environmental risk factors induced chronic inflammation and α-synuclein aggregation in the gut of rats

doi: 10.1038/s41531-026-01281-3

Figure Lengend Snippet: LRRK2 R1627P mutation significantly reduced intestianl endogenous total LRRK2, pT73-Rab10 protein level and the number of Paneth cells. Compared with age-macthed WT rats, LRRK2 R1627P rats showed shorter small intestine length, villus height, and crypt depth (from 8 months old); a significant decrease in the number of goblet cells, expression of barrier proteins, and microbial community diversity (from 16 months old); and a marked increase in the TLR4/MyD88/NF-κB signaling pathway, the number of M1 macrophages, the expression of p S129 -α-Syn, and the abundance of Lactobacillus (from 16 months old). Long-term administration of the TLR4 inhibitor TAK-242 effectively ameliorated the imbalance in intestinal homeostasis of LRRK2 R1627P rats.

Article Snippet: Antibody information: LRRK2 (Abcam, ab133474, 1:200), βIII-Tubulin (Servicebio, GB15139, 1:500), CD68 (Santa Cruz Biotechnology, sc-20060, 1:100), CD163 (Servicebio, GB11340-1, 1:200), iNOS (Affinity Biosciences, AF0199, 1:200), IL-6 (Cell Signaling Technology, D5W4V, 1;200), p S129 -α-Syn (Cell Signaling Technology, 23706S, 1:200).

Techniques: Mutagenesis, Expressing

Journal: bioRxiv

Article Title: Mitoxantrone Hydrochloride Targets APP and LRRK2 to Improve Neurodegeneration in Parkinson’s Models

doi: 10.1101/2025.10.06.680832

Figure Lengend Snippet: 6-OHDA (4 μg) was stereotaxically injected into 4-month-old C57 mice on the left striatum (day 0), and MH was administered via i.p. (0.5 mg/kg) or oral gavage (1 mg/kg) on day 8 for 5 days (cumulative doses of i.p. 2.5 mg/kg and oral 5 mg/kg respectively) before behavioral test after 2-, 6-, and 12-weeks of MH treatment. ( A-D ). 2-, 6-, and 12-weeks after MH treatment, anti-clockwise ipsilateral rotation in the Rotation Test ( A ), left rearing ratio in the Cylinder Test ( B ), riding duration in the Rotarod Test ( C ), and time to descend the pole in the Pole Test ( D ) were plotted. For each time point, animal behavioural was normalized to the vehicle control. Data are presented as the mean ± SD; n=4 per group. * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001 by one-way ANOVA with Tukey’s post hoc test. ( E ) After performing behavioural tests 12-weeks post drug treatment, mice were sacrificed. The striatum tissues from left side lesion (L) and intact right side (R) of the mouse brain were used for western blot analysis. ( F-J ) Quantification of LRRK2 ( F ), APP ( G ), TH ( H ), pS 129 -α-synuclein to α-synuclein ( I ), and pS 199/202 -Tau ( J ) from western blot in E . Protein levels are presented as the left to right ratio and normalized to the vehicle group. Data are presented as the mean ± SD; n=3-6 per group. * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001 by one-way ANOVA with Tukey’s post hoc test. ( K ) After performing behavioural tests 12-weeks post drug treatment, mice were sacrificed. The mouse brains were immunohistochemical stained with TH. The dash line area represents the SNpc region. Scale bar = 100 μm. ( L ) Quantification of integrative TH+ DA neuron density in K . Data are presented as the mean ± SD; n=3. ** P < 0.01, *** P < 0.001, and **** P < 0.0001 by two-way ANOVA with multiple comparison.

Article Snippet: The primary antibodies used in this study were LRRK2 (Abcam, #ab133474), APP, LC3II (Abcam, #ab243506), tyrosine hydroxylase (TH) (Millipore, #MAB318), p62 (Cell Signaling, #39749,), p-α-synuclein (S129) (Cell Signaling, #23706S), cleaved caspase 3 (Cell Signaling, #9661S), pS199/202-Tau (Milipore, #ab9674), α-synuclein (BD Transduction Lab, #610787), pThr 73 Rab10 (Abcam, #ab230261), Rab 10 (Cell Signaling, #8127,) and β-actin (Santa Cruz, #AC-15).

Techniques: Injection, Control, Western Blot, Immunohistochemical staining, Staining, Comparison

Journal: iScience

Article Title: Distinct microRNA signatures define sporadic PSP-RS and PD in patient-derived midbrain organoids

doi: 10.1016/j.isci.2025.113162

Figure Lengend Snippet: PD-derived MOs recapitulate hallmark pathological features (A) Western blot analysis at day 90 reveals increased levels of phosphorylated α-synuclein at Ser129 (pS129) in PD midbrain organoids (MOs) compared to healthy controls (HC). GAPDH was used as a loading control. (B) Densitometric quantification of pS129-α-synuclein levels normalized to GAPDH. Data are presented as mean ± SEM ( n = 3). p < 0.05, Welch’s t test. (C) Immunoblot showing elevated levels of oligomeric and tetrameric α-synuclein in PD MOs relative to controls. Molecular weights are indicated for α-synuclein multimers. (D) Quantification of oligomeric α-synuclein signal intensity normalized to GAPDH. Data represent mean ± SEM ( n = 3). ∗∗ p < 0.001, Welch’s t test. (E) Immunofluorescence analysis of PD MO cryosections stained for MAP2 and pS129-α-synuclein reveals intracellular inclusions resembling Lewy body-like structures (white arrowhead, enlarged in right panel). Nuclei were counterstained with DAPI. Scale bars, 50 μm; 10 μm higher magnification. (F) Immunostaining for tyrosine hydroxylase (TH) and quantification of mean fluorescence intensity (MFI) show reduced dopaminergic marker expression in PD MOs compared to HC. Data are presented as mean ± SEM from at least 12 regions of interest (ROIs) across 3 biological replicates. p < 0.05, Welch’s t test. Scale bars, 25 μm.

Article Snippet: p-α-synuclein (Ser129) (1:500) , Cell Signaling Technology , Cat#23706: RRID:AB_2798868.

Techniques: Derivative Assay, Western Blot, Control, Immunofluorescence, Staining, Immunostaining, Fluorescence, Marker, Expressing