mouse anti sox2 primary antibody  (Santa Cruz Biotechnology)

Journal: bioRxiv

Article Title: White and Clearing: New Optical Tissue Clearing Method

doi: 10.1101/2025.01.30.635653

Figure Lengend Snippet: Effect of TDE incubation in endogenous TdTomato and EGFP fluorescence in LysMCre +/− , mT/mG mouse heart samples and AlbCre +/− , mT/mG mouse liver samples. A, B, C, D) % of the original Signal-to-noise/background ratio values over 24h for TdTomato in samples incubated in PBS (control), TDE 60%, 80%, 90%, and 100% plus additional post- incubation PBS wash for A) TdTomato and B) EGFP in heart samples, C) TdTomato and D) EGFP in liver samples. N=3. Error bars shown as standard error. E) Representative images of TdTomato (left, red) and EGFP (right, green) visualization in liver samples after 24h incubation in PBS, 60% TDE 80% TDE, 90% TDE and TDE 100%. Confocal microscopy images, 10x magnification. F,G,H) Effect of TDE incubation in AlexaFluor 488 fluorescence. F) Signal-to- noise ratio values over time for AlexaFluor 488 in samples incubated in PBS (control) and TDE 97%. G,H) examples of DAPI (Blue) and AlexaFluor 488 (Green) visualization in samples incubated for 1h in B) PBS (control) and C) TDE 97%. 20x Confocal Microscopy images. Scale bar = 37.2µm. I,J,K) Results of DAPI (Blue) and Sox2 (Green) IHC labelling in R1+H2O2+TDE- cleared mouse brain samples. 10x magnification confocal microscopy. I) DAPI, B) Sox2, K) channel merge. Bleaching: 45min 10% H2O2 in PBS, 65°C, glass container. R1+ TDE: 10 days R1 + 1-day TDE 97%, 37°C, shaker.

Article Snippet: For the immunohistochemistry-immunofluorescence (IHC-IF) assays, thin-slice samples were incubated in 1 µg/mL mouse anti-Sox2 primary antibody (sc-365823, Santa Cruz Biotechnology, TX, USA) in a shaker at 4°C overnight.

Techniques: Incubation, Fluorescence, Control, Confocal Microscopy

Journal: iScience

Article Title: Generation and characterization of inducible KRAB-dCas9 iPSCs from primates for cross-species CRISPRi

doi: 10.1016/j.isci.2024.110090

Figure Lengend Snippet: Efficient SOX2 knockdown can be induced in the KRAB-dCas9 iPSCs (A) Phase contrast images of human H.i2_clone2, gorilla G.i1_clone3, and cynomolgus C.i2_clone1. KRAB-dCas9 iPSCs were either transduced with a SOX2 -targeting gRNA or remained non-transduced and were cultured in medium with or without 1 μg/mL dox for 4 days, respectively. Close-up of the cells that were transduced with a SOX2 -targeting gRNA and cultured under dox-conditions; scale bar represents 500 μm. (B) SOX2 transcripts of perturbed or unperturbed cells were quantified by qPCR for the three human, three gorilla and two cynomolgus KRAB-dCas9 iPS cell lines. Expression levels were normalized to GAPDH and relative SOX2 transcript expression between dox-treated and untreated cells was determined using the ΔΔCt method. Negative ΔΔCt-values comparing the +dox and ˗dox condition are shown for SOX2 . Horizontal lines indicate the mean; ( n = 3 technical replicates). Results of paired t tests indicate differences between the mean of the samples with and without a SOX2 -targeting gRNA (0 '∗∗∗∗' 0.0001 '∗∗∗' 0.001 '∗∗' 0.01 '∗' 0.05 'ns' Inf). A t test across species between all samples with and all samples without a SOX2 -targeting gRNA revealed a significant difference of the SOX2 ΔΔCt ( p value = 4.5 x 10 −8 ). (C) Negative ΔΔCt of SOX2 in perturbed cells compared to negative ΔCt of KRAB-dCas9. Data are represented as mean ± SD; Pearson’s r = −0.53, p value = 0.174.

Article Snippet: Furthermore, first SOX2 was detected with an anti-SOX2 primary antibody (1:1,000; Cell Signaling Technologies, 4900S; RRID: AB_10560516 ) and an HRP-coupled anti-mouse secondary antibody.

Techniques: Knockdown, Transduction, Cell Culture, Expressing

Journal: iScience

Article Title: Generation and characterization of inducible KRAB-dCas9 iPSCs from primates for cross-species CRISPRi

doi: 10.1016/j.isci.2024.110090

Figure Lengend Snippet: SOX2 knockdown in the KRAB-dCas9 iPSCs leads to reduced association with a pluripotent cell profile (A) Immunofluorescence stainings of SOX2 in human H.i2_clone2, gorilla G.i1_clone3, and cynomolgus C.i2_clone1 cells. KRAB-dCas9 iPSCs with an integrated SOX2-targeting gRNA were cultured in medium with or without 1 μg/mL dox for 4 days; scale bars indicate 250 μm. (B) Western blot analysis for SOX2 and beta-actin of human, gorilla, and cynomolgus KRAB-dCas9 iPSC clones with an integrated SOX2-targeting gRNA. Cells were cultured in medium with or without 1 μg/mL dox for 4 days, before protein extraction. (C) log2FC of the SOX2 expression level in KRAB-dCas9 iPSCs with an integrated SOX2 -targeting gRNA between +dox and ˗dox condition; error bars indicate SEM; (∗∗∗∗ ( p .adj ≤ 0.0001), ns ( p .adj > 0.05)). (D) Cell type classification was performed using SingleR with reference data from Rhodes et al. (see also Figure S4 ). Correlation scores of the samples to pluripotent cells of the reference data are shown for the +dox and ˗dox condition; results of paired t tests indicate significant differences of the mean scores between the two conditions (0 '∗∗∗∗' 0.0001 '∗∗∗' 0.001 '∗∗' 0.01 '∗' 0.05 'ns' Inf); note that the reference data were generated from human samples only and hence lower correlation scores are expected for more diverged primates. (E) Correlation of the Δscore for pluripotent cells between the +dox and the ˗dox score to the log2FC of SOX2 expression; data are represented as mean (n = 4–5 biological replicates) +/− SEM; Pearson’s r = 0.91, p value = 0.002.

Article Snippet: Furthermore, first SOX2 was detected with an anti-SOX2 primary antibody (1:1,000; Cell Signaling Technologies, 4900S; RRID: AB_10560516 ) and an HRP-coupled anti-mouse secondary antibody.

Techniques: Knockdown, Immunofluorescence, Cell Culture, Western Blot, Clone Assay, Protein Extraction, Expressing, Generated

Journal: iScience

Article Title: Generation and characterization of inducible KRAB-dCas9 iPSCs from primates for cross-species CRISPRi

doi: 10.1016/j.isci.2024.110090

Figure Lengend Snippet:

Article Snippet: Furthermore, first SOX2 was detected with an anti-SOX2 primary antibody (1:1,000; Cell Signaling Technologies, 4900S; RRID: AB_10560516 ) and an HRP-coupled anti-mouse secondary antibody.

Techniques: Virus, Recombinant, Membrane, Lysis, CRISPR, DNA Extraction, Western Blot, Transfection, Reverse Transcription, Bicinchoninic Acid Protein Assay, SYBR Green Assay, Stripping, Software, Microscopy

Journal: Genes

Article Title: Role of Neurocellular Endoplasmic Reticulum Stress Response in Alzheimer’s Disease and Related Dementias Risk

doi: 10.3390/genes15050569

Figure Lengend Snippet: Validation of generated NSCs. ( a ) A schematic outline of the NSC differentiation method. ( b ) Representative ICC images showing expression of NSC markers Nestin, PAX6, SOX2, and SOX1 in generated NSCs. ( c ) Heap map of transcriptome-wide differentially expressed genes between iPSCs and their differentiated NSCs. ( d ) Cell type gene set (PanglaoDB Augmented 2021) enrichment analysis plot of the generated NSCs’ upregulated transcriptome. ( e ) Bar plot showing dorsal neuroepithelial gene expression profile of the generated NSCs.

Article Snippet: The ICC analysis of the NSC markers was performed using commercially available the primary antibodies mouse anti-human Nestin (MA1-110, Invitrogen, Thermo Fisher Scientific, Waltham, MA, USA), rabbit anti-human PAX6 (#60443, Cell Signaling Technology, Inc., Danvers, MA, USA), mouse anti-human SOX2 (sc-365964, Santa Cruz Biotechnology, Inc., Dallas, TX, USA), and rabbit anti-human SOX1 (#4194, Cell Signaling Technology, Inc., Danvers, MA, USA); the secondary antibodies donkey anti-mouse Alexa FluorTM 488 and donkey anti-rabbit Alexa FluorTM 594 (R37114 and R37119, respectively; Invitrogen, Thermo Fisher Scientific, Waltham, MA, USA); and standard ICC techniques.

Techniques: Biomarker Discovery, Generated, Expressing, Gene Expression

Journal: Biology Direct

Article Title: ADSC-derived exosomes attenuate myocardial infarction injury by promoting miR-205-mediated cardiac angiogenesis

doi: 10.1186/s13062-023-00361-1

Figure Lengend Snippet: Isolation and characterization of exosomes derived from ADSCs A. The expression of relevant biomarkers of ADSCs (CD29, CD44, CD31 and CD34) were measured by flow cytometry; B. The concentration and size distribution of ADSC-Exo was detected by Nanoparticle tracking analysis (NTA); C. The morphology of ADSC-Exo was characterized by transmission electron microscope; C – E The relative expression of exosomes’ marker proteins (CD63, TSG101 and CD9) evaluated by Western blot;

Article Snippet: All the sections were blocked with 1% goat serum albumin for 1 h and then incubated with mouse monoclonal anti-CD31 primary antibody (Ab955, 1:200; Abcam) at 4 °C overnight.

Techniques: Isolation, Derivative Assay, Expressing, Flow Cytometry, Concentration Assay, Transmission Assay, Microscopy, Marker, Western Blot

Journal: Biology Direct

Article Title: ADSC-derived exosomes attenuate myocardial infarction injury by promoting miR-205-mediated cardiac angiogenesis

doi: 10.1186/s13062-023-00361-1

Figure Lengend Snippet: Intravenous delivery of ADSCs-Exosomes promotes angiogenesis and microvascular endothelial cells proliferation. A. Representative HE staining of neovessels in the hearts from the sham group, MI group and MI + ADSC-exo group; B. The expression of angiogenic marker HIF-1α and VEGF were evaluated by western blot; C. Fluorescent immunostaining of plaque sections with anti-CD31 antibody; D-E. Quantitative analysis of angiogenic marker HIF-1α and VEGF expression. Data were presented as Mean ± SEM, n = 8–10 mice. ** P < 0.05, * P < 0.05

Article Snippet: All the sections were blocked with 1% goat serum albumin for 1 h and then incubated with mouse monoclonal anti-CD31 primary antibody (Ab955, 1:200; Abcam) at 4 °C overnight.

Techniques: Staining, Expressing, Marker, Western Blot, Immunostaining