Celllight late endosomes gfp

Manufactured by Thermo Fisher Scientific

Sourced in United States

The CellLight® Late Endosomes-GFP is a fluorescent labeling tool designed for live-cell imaging of late endosomes in mammalian cells. It utilizes a genetically encoded fluorescent protein (GFP) to enable visualization of late endosomal structures within the cell.

Automatically generated - may contain errors

Lab products found in correlation

5 protocols using celllight late endosomes gfp

Multifunctional Nanoparticles for Drug Delivery

Check if the same lab product or an alternative is used in the 5 most similar protocols

2-cyano-2-propyl benzodithioate (RAFT reagent), poly (ethylene glycol) methacrylate, 2-(Diisopropylamino)ethyl methacrylate, AIBN, tetraethyl orthosilicate (TEOS) and cetyl trimethylammonium bromide (CTAB) were purchased from Sigma Aldrich (Yongin, Korea) and TCI (Japan). Camptothecin was obtained from Ontario Chemicals Inc (Canada) and doxorubicin hydrochloride was obtained from Acorn-Pharma (U.S.A). Fetal bovine serum (FBS) was from Gibco, cell culture medium and reagents were obtained from Invitrogen (Korea). Cell viability analysis was measured using the alamarBlue^® cell viability reagent (DAL 1025, ThermoFisher, Korea) following the manufacturer’s protocol and analyzed using the fluorescence measurement from Tecan – Infinite 200 series reader. CellLight^® Late Endosomes-GFP, BacMam 2.0 (C10588) and LysoTracker^® Green DND-26 (L7526) were obtained from ThermoFisher (Korea). Unless otherwise stated, all used were obtained from commercial suppliers (Sigma Aldrich, TCI and Abcam) and were used as received. DLS measurements were made using a Malvern Nanozetasizer (Nano ZS series). UV-Visible spectra were measure using the JASCO V250 spectrophotometer. The fluorescence spectra were obtained using a JASCO FP-6500 spectrofluorimeter. PEG-PDS polymer was prepared and characterized using the 400 MHz, Bruker AVANCE III HD NMR spectroscopy following our previous literature33 .

Palanikumar L., Jeena M.T., Kim K., Yong Oh J., Kim C., Park M.H, & Ryu J.H. (2017). Spatiotemporally and Sequentially-Controlled Drug Release from Polymer Gatekeeper–Hollow Silica Nanoparticles. Scientific Reports, 7, 46540.

+ Open protocol

+ Expand

Ex4-Loaded Chitosan Nanoparticles for Insulin Delivery

Check if the same lab product or an alternative is used in the 5 most similar protocols

Ex4 was purchased from GL Biochem Ltd. (Shanghai, China). RPMI 1640, fetal bovine serum, rabbit anti-murine and human FATP4 polyclonal antibody (PA5–42446), Alexa Fluor 594–NHS Ester, LysoTracker Green DND-26, CellLight Early Endosomes–GFP, CellLight Late Endosomes–GFP, and ER-Tracker Green were obtained from Thermo Fisher (Waltham, MA, USA). Rabbit anti-murine insulin antibody was from Abcam (ab63820, Cambridge, MA, USA). Chitosan (molecular weight, 50,000 to 190,000 Da), pentasodium triphosphate, zinc trifluoroacetate hydrate, 16-hydroxyhexadecanoic acid, silver enhancer kit, and diisopropyl ether were ordered from Sigma-Aldrich (St. Louis, MO, USA). Mono-Sulfo-NHS-Nanogold (1.4 nm) was purchased from Nanoprobe, Inc. (Yaphank, NY, USA), and 1,2-dioleoyl-sn-glycero-3-phosphate (sodium salt) and cholesterol were purchased from Avanti Polar Lipids Inc. (Alabaster, AL, USA).

Hu Z., Nizzero S., Goel S., Hinkle L.E., Wu X., Li C., Ferrari M, & Shen H. (2020). Molecular targeting of FATP4 transporter for oral delivery of therapeutic peptide. Science Advances, 6(14), eaba0145.

+ Open protocol

+ Expand

Nanoparticle-mediated Cell Viability Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols

The following reagents were used: horse heart Cyt c (C7752, Sigma-Aldrich), EDC (E6383, Sigma-Aldrich), NHS (130672, Sigma-Aldrich), 2-(N-morpholino)ethane sulfonic acid hydrate, 4-morpholineethanesulfonic acid (M8250, Sigma-Aldrich), l-ascorbic acid (A92902, Sigma-Aldrich), μ-plate 24-well black ibiTreat surface (IB-82426, Thistle Scientific), Hoechst 33342 (NucBlue Live ReadyProbes Reagent, R37605, Thermo Fisher Scientific), actin using Phalloidin–iFluor 488 conjugate (23115, AAT Bioquest, Stratech), PrestoBlue HS cell viability reagent (P50200, Invitrogen), calcein AM (C1430, Thermo Fisher Scientific), propidium iodide (P4170, Sigma-Aldrich), H2DCFDA (D399, Invitrogen), CellEvent Caspase-3/7 Green Flow Cytometry Assay Kit (C10427, Invitrogen), Zombie NIR Fixable Viability Kit (423105, BioLegend), CellEvent Caspase-3/7 Green ReadyProbes Reagent (R37111, Invitrogen), CellLight Late Endosomes-GFP, BacMam 2.0 (C10588, Thermo Fisher Scientific) and LysoTracker Green DND-26 (L7526, Thermo Fisher Scientific). A catalogue number is not available for the carboxylic-PEG-coated GNPs or Z as they were customized and purchased from Nanopartz and Porphychem, respectively.

Jain A., Gosling J., Liu S., Wang H., Stone E.M., Chakraborty S., Jayaraman P.S., Smith S., Amabilino D.B., Fromhold M., Long Y.T., Pérez-García L., Turyanska L., Rahman R, & Rawson F.J. (2023). Wireless electrical–molecular quantum signalling for cancer cell apoptosis. Nature Nanotechnology, 19(1), 106-114.

+ Open protocol

+ Expand

Visualizing endosome dynamics in fibroblasts

Check if the same lab product or an alternative is used in the 5 most similar protocols

Fibroblasts were grown on a sterile tissue culture dish with a cover glass bottom (FD35-100, FluoroDish, World Precision Instruments) and were cotransfected with the RFP-Rab5a–expressing vector and the GFP-Rab7a–expressing vector (CellLight Late Endosomes-GFP, Life Technologies). Live cells were imaged 48 hours after transfection using a Zeiss LSM 710 confocal microscope (Carl Zeiss) and analyzed using Zeiss Zen 2009 and ImageJ (NIH). For quantitative studies of early endosome volume, fibroblasts were cotransfected with the RFP-Rab5a vector as above, and after 48 hours culture medium was removed and cells were washed with PBS. Cells were fixed for 20 minutes with 4% paraformaldehyde in PBS at room temperature, and then washed 3 times with PBS. Ten images/cells per patient and control were collected using a Zeiss LSM 710 confocal microscope (Carl Zeiss). Deconvolution of raw images was carried out using Huygens Software (Scientific Volume Imaging); endosome volume was measured using Imaris Software (Bitplane). Statistical analysis was performed in Prism (GraphPad), using 1-way ANOVA and Tukey’s post hoc test (P < 0.05). These studies were done in a blind fashion by the investigator.

Buonocore F., Kühnen P., Suntharalingham J.P., Del Valle I., Digweed M., Stachelscheid H., Khajavi N., Didi M., Brady A.F., Blankenstein O., Procter A.M., Dimitri P., Wales J.K., Ghirri P., Knöbl D., Strahm B., Erlacher M., Wlodarski M.W., Chen W., Kokai G.K., Anderson G., Morrogh D., Moulding D.A., McKee S.A., Niemeyer C.M., Grüters A, & Achermann J.C. (2017). Somatic mutations and progressive monosomy modify SAMD9-related phenotypes in humans. The Journal of Clinical Investigation, 127(5), 1700-1713.

+ Open protocol

+ Expand

Intracellular Nanoparticle Tracking in Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

Confocal images where obtained using a Leica SP8 CLSM with a 63 × 1.2 water objective. For NR668 excitation, a white light laser at 514 nm was used, and emission was detected at 580–660 nm using a photon counting hybrid detection system. Z-stacks of cells were obtained to verify that the NPs were intracellular.
In PC3 cells, lysosomes were stained with the pH sensitive dye LysoTracker Blue (DND-22, Life Technologies) at 2.5 μM for 1 h and imaged using a pulsed multiphoton laser at 780 nm to excite the dye. In RBE4 cells early endosomes, late endosomes and lysosomes were labeled by cellular transduction using CellLight Early Endosomes-GFP, CellLight Late Endosomes-GFP and CellLight Lysosomes-GFP (Life Technologies), respectively, at a concentration of 40 particles per cell 24 h prior to imaging. After labeling the organelles, cells were incubated with NPs for 3 h before imaging.

Sulheim E., Baghirov H., von Haartman E., Bøe A., Åslund A.K., Mørch Y, & Davies C.D. (2016). Cellular uptake and intracellular degradation of poly(alkyl cyanoacrylate) nanoparticles. Journal of Nanobiotechnology, 14, 1.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!