anti class iii betatubulin tubb3 antibody  (R&D Systems)

Journal: Cellular and Molecular Neurobiology

Article Title: Diverse Effects of Various Toll-Like Receptor 2 Ligands on Neuronal Activity and Cell Death

doi: 10.1007/s10571-025-01632-3

Figure Lengend Snippet: Expression of TLR2 in the cortical-derived neurons and other cells. Primary rat cortical cells were cultured for 2–16 days. ( a ) Representative images of primary cortical cells at DIV2, DIV9, DIV12, and DIV16. Representative images showing the signal from Hoechst 33342-stained nuclei in blue, beta III tubulin immunoreactivity in green, and TLR2 immunoreactivity in red in the cultured cortical cells, and merged image (bottom panel). (4 biological replicates in 1 technical replicate from 2 independent experiments) ( b ) Magnified images from ( a ) showing TLR2 in neurons. Scale bar: 100 μm. Cells were analyzed in 4 fields of view. Two independent experiments were performed

Article Snippet: Anti-class III beta-tubulin (TUBB3) antibody (R&D systems, MAB1195), a marker for neurons (Ballas et al. ), anti-TLR2 antibody (Thermo, MA532787 ), anti-glial fibrillary acidic protein (GFAP) antibody (abcam, ab68428), a marker for astrocytes (Shixing et al. ), anti-Iba1 antibody (CST, 17198T), a marker for microglia (Ni et al. ), and anti-Olig2 antibody (abcam, ab109186), a marker for oligodendrocytes (Xu et al. ), were used as primary antibodies (Wyczanska et al. ).

Techniques: Expressing, Derivative Assay, Cell Culture, Staining

Journal: Materials Today Bio

Article Title: Dorsal root ganglion-targeted analgesic delivery for effective relief of neuropathic pain

doi: 10.1016/j.mtbio.2025.102025

Figure Lengend Snippet: In vivo DRG-targeting ability of BK-LNPs in SNI and CINP. (A) t-distributed stochastic neighbor embedding (t-SNE) visualization of Adam8 mRNA expression patterns across DRG cellular subpopulations and (B) RNAscope imaging for Adam8 mRNA in the DRG neurons following SNI modeling. (C–D) Relative Adam8 mRNA expression level in the DRG on the 7th and 28th days following SNI modeling. (E) CLSM images of the bilateral DRGs ipsilateral and contralateral to SNI surgical site and (F) corresponding quantitative analysis of RhB fluorescence. Tuj1 was a neuronal marker. (G) Schematic illustration for the bilateral DRGs ipsilateral and contralateral to SNI surgical site. (H) Relative Adam8 mRNA expression level in the DRG as well as in the heart, liver, spleen, lung, and kidney on the 7th day following CINP modeling. (I) CLSM images of the DRG in CINP and (J) corresponding quantitative analysis of RhB fluorescence. For CLSM imaging, mice were intravenously injected with LNP/RhB and BK-LNPs/RhB and at 6 h post-injection, mice were euthanized for analysis. Data were expressed as mean ± SEM (n = 3 for C and D; n = 5 for F, H, and J). ∗∗ p < 0.01 and ∗∗∗ p < 0.001.

Article Snippet: The obtained DRG sections (14 μm in thickness) were blocked with 10 % BSA for 1 h and then sequentially incubated with neuronal class III β-tubulin (Tuj1) primary antibody (D71G9, rabbit, Cell Signaling Technology, #5568, 1:1000) overnight and its fluorescence-labeled secondary antibody (Alexa Fluor 488 anti-rabbit IgG, donkey, ThermoFisher, #A-21206, 1:500) for 2 h. The fluorescence of Tuj1 and RhB was captured on the SP8 CLSM using the corresponding filters to evaluate the targeting ability of BK-1361.

Techniques: In Vivo, Expressing, RNAscope, Imaging, Fluorescence, Marker, Injection

Journal: Scientific Reports

Article Title: An optimized method for directed differentiation of hypothalamic neural stem cells in a 3D culture system

doi: 10.1038/s41598-025-02847-6

Figure Lengend Snippet: Validation of htNSCs differentiation into GnRH-like neurons within the Matrigel-based 3D culture system. ( A ) Differentiation of individual htNSCs into neuron-like cells; days 0–4: induction medium I; days 4–25: induction medium II. ( B – E ) Immunostaining for Tuj1 (red) and GnRH (green) in differentiated htNSCs cultured in the Matrigel-based 3D system at day 11. ( F – M ) Immunostaining for Map2 (red) and GnRH (green) in differentiated htNSCs cultured in the Matrigel-based 3D system at day 18 and day 25. ( B – M ) Scale bar: 100 μm. ( N ) qPCR analysis of Gnrh1 mRNA expression in cells and ( O ) ELISA analysis of GnRH protein levels in supernatants collected at different time points during htNSCs and ctNSCs differentiation in the Matrigel-based 3D system. Data are presented as mean ± SEM and analyzed using unpaired Student’s t -tests. **** P < 0.0001, *** P < 0.001, ** P < 0.01, * P < 0.05, and “ns” for no significant difference, for n = 3 independent experiments. htNSCs, hypothalamic neural stem cells; Tuj1, neuron-specific class III beta-tubulin; Map2, microtubule-associated protein 2; GnRH, gonadotrophin-releasing hormone; DAPI, 4’,6-diamidino-2-phenylindole.

Article Snippet: Neuronal Class III β-Tubulin (Tuj1) , Cell Signaling Techology , 4466 S , Mouse/1:200.

Techniques: Biomarker Discovery, Immunostaining, Cell Culture, Expressing, Enzyme-linked Immunosorbent Assay