Anti GFP  
DyLight (商標名)

Western Bot of Goat Anti-GFP (GOAT) Antibody DyLight^TM 488 Conjugated. Lane 1: Opal Prestained Molecular Weight Marker (p/n MB-210-0500). Lane 2: GFP / HEK293T Whole Cell Lysate Reduced [0.05/10μg]. Lane 3: HEK293T Whole Cell Lysate Reduced (p/n W09-001-GX5) [10μg]. Lane 4: GFP / NIH-3T3 Whole Cell Lysate Reduced [0.05/10μg]. Lane 5: NIH-3T3 Whole Cell Lysate Reduced (p/n W10-000-358) [10μg]. Lane 6: GFP / PC-12 Whole Cell Lysate Reduced [0.05/10μg].Lane 7: PC-12 Whole Cell Lysate Reduced (p/n W12-001-GL9) [10μg]. Secondary Antibody: Goat Anti-GFP (GOAT) Antibody DyLight^TM 488 Conjugated at 1.0μg/mL overnight at 2-8°C. Blocking Buffer: Blocking Buffer for Fluorescent Western Blotting (p/n MB-070) for 1 hour at RT. Expect: 27kDa.

Western Blot of DyLight^TM 488 conjugated anti-GFP antibody to detect GFP control proteins. Lane 1: His-Sumo-GFP. Lane: Molecular Weight. Lane 2: Beta-Galactosidase (negative control). Lane 3: recombinant GFP control protein (000-001-215). Load: 35 μg per lane. Primary antibody: none. Secondary antibody: DyLight^TM 488 conjugated anti-GFP goat secondary antibody at 1:5,000. Block: MB-070 for 2 hr at RT. Predicted/Observed size: 27kDa/54kDa, 27kDa for rGFP/~45kDa His-Sumo-GFP. Other band(s): none.

P2X7-Rs are highly expressed in ON-RGCs of Thy1-YFP-H mice. Thy1-YFP-H RGCs (green) are projections of the original confocal Z-stacks, while the P2X7-R puncta contacting the dendrites (red) are digitally rendered as spots (presented as 3 times larger than the true colocalised puncta for ease of viewing). (A？C) Thy1-YFP-H labelled ON-RGC (green) showing dense labelling for P2X7-R puncta (red) shown in (A) orthogonal view, (B) from above and (C) magnified inset from (B). (D) A single plane of raw data from the ON-cell (green) with P2X7-R staining (red). (E？G) Thy1-YFP-H labelled OFF-RGC (green) showing sparser labelling for P2X7-R puncta (red) than ON-RGCs. The OFF-RGC is shown in (E) orthogonal view, (F) from above and (G) magnified inset from？F. (H？L) Thy1-YFP-H labelled ON？OFF RGC (green) in the orthogonal view (H) displaying dendritic fields stratifying in ON and OFF sublaminae. Labelling for the P2X7-R puncta (red) in the ON sublamina (I) from above and (J) magnified inset from (I). Labelling for the P2X7-R puncta (red) in the OFF sublamina (K) from above and (L) magnified inset from K. (M) Quantification of P2X7-R puncta in the ON and OFF lamina of the IPL for ON-, OFF- and ON？OFF RGCs. Numbers indicate number of cells from 4 animals. *p？<？0.01; one-way ANOVA, Tukey’s multiple comparisons test. Scale bar is 10？μm.Fig 6. PMID:？33603067

(C) Passive diffusion from the cytoplasm to the nucleus measured by FRAP, in HeLa cells transduced with mTagBFP-2A control, SUN1 or SUN2 lentivectors and a lentivector encoding GFP. Top, representative layout of a photobleached control HeLa cell: imaging starts at t = 0 after high intensity laser exposure and ends at t = 80 seconds, when fluorescence in the nucleus has been recovered (scale bar: 5 μm). Bottom left: GFP intensity recovery over time (s). For each individual cell, the background intensity was subtracted and intensity was normalized to 0 at t0 after photobleaching while the max intensity reached during the course of each measurement was set to 1. Curves show mean and standard deviation per time point across indicated number of cells per condition. Bottom right, hillslopes for each cell were calculated via non-linear regression fit (total number of nuclei analyzed per cell line are indicated, n=1 experiment; un-paired ANOVA one-way with Dunnett’s post-test, line at mean ± SEM). FS5. PMID:？34592156

Properties of DyLight^TM Fluorescent Dyes. The emission spectra for this DyLight^TM conjugate match the principle output wavelengths of most common fluorescence instrumentation.

Comparison of fluorescence absorbtion and emission spectra for DyLight^TM 488 and Alexa Fluor 488 in PBS, pH7.2. The emission spectra for this DyLight^TM conjugate match the principle output wavelengths of most common fluorescence instrumentation.

Multi-color Immunofluorescence Microscopy using DyLight^TM dyes. Tissue: human breast carcinoma. Fixation: 0.5% PFA. Antigen retrieval: not required. Primary antibody: Anti-Histone and Anti-Tubulin antibody at 10 μg/mL for 1 h at RT. Secondary antibody: DyLight^TM 488 conjugate and DyLight^TM 549 conjugate goat secondary antibody at 1:10,000 for 45 min at RT. Localization: Histone is nuclear and Tubulin is cytoplasmic. Staining: Anti-Histone detection using a DyLight^TM 488 conjugate (green) fluorescent signal and Anti-Tubulin was detected using a DyLight^TM 549 conjugate (red) fluorescent signal. Nuclei were counter-stained using DAPI (blue).