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A ) Rat primary cortical neurons were co-transfected at DIV3 with YFP-ArfGAP1 (WT, 3A or 3D) or empty <t>vector</t> <t>and</t> <t>DsRed-Max-N1</t> plasmids, and fixed at DIV6 for confocal fluorescence microscopy analysis. Fluorescent images reveal the co-labeling of cortical neurons with YFP-ArfGAP1 (green) and DsRed (red), with the DsRed images pseudo colored (ICA) to enhance the contrast of neuritic processes. Neuronal soma (white arrows) and axonal processes (black arrowheads) are indicated. Scale bars: 50 μm. B ) Quantitative analysis of DsRed-positive axon length in YFP-ArfGAP1-positive neurons or control neurons (empty vector) is shown. Bars represent the mean ± SEM axon length (in μm) from 90-120 double DsRed-/YFP-ArfGAP1-positive neurons, or single DsRed-positive neurons (control), across at least three independent experiments/cultures. *** P <0.001 or **** P <0.0001 compared to control (DsRed alone), or as indicated, by one-way ANOVA with Dunnett’s multiple comparisons test. ns , non-significant.
Dsred Max N1, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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A ) Rat primary cortical neurons were co-transfected at DIV3 with YFP-ArfGAP1 (WT, 3A or 3D) or empty vector and DsRed-Max-N1 plasmids, and fixed at DIV6 for confocal fluorescence microscopy analysis. Fluorescent images reveal the co-labeling of cortical neurons with YFP-ArfGAP1 (green) and DsRed (red), with the DsRed images pseudo colored (ICA) to enhance the contrast of neuritic processes. Neuronal soma (white arrows) and axonal processes (black arrowheads) are indicated. Scale bars: 50 μm. B ) Quantitative analysis of DsRed-positive axon length in YFP-ArfGAP1-positive neurons or control neurons (empty vector) is shown. Bars represent the mean ± SEM axon length (in μm) from 90-120 double DsRed-/YFP-ArfGAP1-positive neurons, or single DsRed-positive neurons (control), across at least three independent experiments/cultures. *** P <0.001 or **** P <0.0001 compared to control (DsRed alone), or as indicated, by one-way ANOVA with Dunnett’s multiple comparisons test. ns , non-significant.

Journal: bioRxiv

Article Title: LRRK2 regulates ArfGAP1 membrane localization, activity and neuronal toxicity via phosphorylation within its lipid-sensing ALPS2 motif

doi: 10.64898/2026.01.12.699049

Figure Lengend Snippet: A ) Rat primary cortical neurons were co-transfected at DIV3 with YFP-ArfGAP1 (WT, 3A or 3D) or empty vector and DsRed-Max-N1 plasmids, and fixed at DIV6 for confocal fluorescence microscopy analysis. Fluorescent images reveal the co-labeling of cortical neurons with YFP-ArfGAP1 (green) and DsRed (red), with the DsRed images pseudo colored (ICA) to enhance the contrast of neuritic processes. Neuronal soma (white arrows) and axonal processes (black arrowheads) are indicated. Scale bars: 50 μm. B ) Quantitative analysis of DsRed-positive axon length in YFP-ArfGAP1-positive neurons or control neurons (empty vector) is shown. Bars represent the mean ± SEM axon length (in μm) from 90-120 double DsRed-/YFP-ArfGAP1-positive neurons, or single DsRed-positive neurons (control), across at least three independent experiments/cultures. *** P <0.001 or **** P <0.0001 compared to control (DsRed alone), or as indicated, by one-way ANOVA with Dunnett’s multiple comparisons test. ns , non-significant.

Article Snippet: A plasmid containing DsRed-Max-N1 was obtained from Addgene (#21718) [ ].

Techniques: Transfection, Plasmid Preparation, Fluorescence, Microscopy, Labeling, Control

A ) Rat primary cortical neurons were co-transfected at DIV3 with YFP-ArfGAP1 (WT, 3A or 3D), FLAG-LRRK2 (G2019S) and DsRed-Max-N1 plasmids, and fixed at DIV6 for immunofluorescence analysis with anti-FLAG antibody. Fluorescent images reveal the co-labeling of cortical neurons with YFP-ArfGAP1 (green), G2019S LRRK2 (yellow) and DsRed (red), with the DsRed images pseudo colored (ICA) to enhance the contrast of neuritic processes. Neuronal soma (white arrows) and axonal processes (black arrowheads) are indicated. Scale bars: 50 μm. B ) Quantitative analysis of DsRed-positive axon length in YFP-ArfGAP1-/LRRK2-positive neurons is shown. Bars represent the mean ± SEM axon length (in μm) from 90-120 triple DsRed-/YFP-ArfGAP1-/LRRK2-positive neurons across at least three independent experiments/cultures. **** P <0.0001 compared to WT ArfGAP1/LRRK2 by one-way ANOVA with Dunnett’s multiple comparisons test. ns , non-significant.

Journal: bioRxiv

Article Title: LRRK2 regulates ArfGAP1 membrane localization, activity and neuronal toxicity via phosphorylation within its lipid-sensing ALPS2 motif

doi: 10.64898/2026.01.12.699049

Figure Lengend Snippet: A ) Rat primary cortical neurons were co-transfected at DIV3 with YFP-ArfGAP1 (WT, 3A or 3D), FLAG-LRRK2 (G2019S) and DsRed-Max-N1 plasmids, and fixed at DIV6 for immunofluorescence analysis with anti-FLAG antibody. Fluorescent images reveal the co-labeling of cortical neurons with YFP-ArfGAP1 (green), G2019S LRRK2 (yellow) and DsRed (red), with the DsRed images pseudo colored (ICA) to enhance the contrast of neuritic processes. Neuronal soma (white arrows) and axonal processes (black arrowheads) are indicated. Scale bars: 50 μm. B ) Quantitative analysis of DsRed-positive axon length in YFP-ArfGAP1-/LRRK2-positive neurons is shown. Bars represent the mean ± SEM axon length (in μm) from 90-120 triple DsRed-/YFP-ArfGAP1-/LRRK2-positive neurons across at least three independent experiments/cultures. **** P <0.0001 compared to WT ArfGAP1/LRRK2 by one-way ANOVA with Dunnett’s multiple comparisons test. ns , non-significant.

Article Snippet: A plasmid containing DsRed-Max-N1 was obtained from Addgene (#21718) [ ].

Techniques: Transfection, Immunofluorescence, Labeling