Organelle Lights™ reagents
The illumination you need, the ease you appreciate:
-
- Free yourself from making your own constructs–try our well characterized and easy to use fluorescent protein-based reagents
- Multiplex easily with other Organelle Lights™ constructs, fluorescent organic dyes, or Qdot® conjugates
- Study dynamic cellular processes and use for subcellular co-localization experiments with precise spatial and temporal resolution
- Get efficient, non-cytopathic delivery to multiple cell types (including primary and stem cells) by BacMam delivery
 |
| | | |||
|
 Figure 1.
|
Table 1
Expression tag details  | ||||
| Organelle Lights™ | CFP | GFP | OFP | RFP |
| (440/480) | (488/510) | (548/565) | (555/584) | |
| Cytoplasm | O36227 | |||
| Endoplasmic Reticulum | O36212 | O36223 | O36230 | |
| Endosomes | O10104 | O36231 | ||
| Golgi | O36215 | O36224 | O10098 | |
| Lysosomes | O36228 | O10100 | ||
| Mitochondria | O36210 | O36222 | O36229 | |
| Nuclear Envelope | O36213 | |||
| Nucleus | O36218 | O36209 | O10099 | |
| Peroxisome | O36211 | |||
| Plasma Membrane | O36216 | O36214 | O36226 | O10139 |
| Synaptophysin | C10080 | |||
| Null virus (control) | C10130 | |||
| Note: Different cell types will have different transfection efficiencies with baculovirus -based technologies. Certain cell types, such as macrophages and other hematopoiteic cells, cannot be transfected with baculovirus. | ||||
|
| Cell Types Transduced using BacMam Technology | Reference / Lab |
|---|---|
| Primates | |
| Human cells | |
| 143TK- | 10 |
| Astroglioma U373MG | 28 |
| Bone marrow fibroblasts | 5 |
| CHP212 (neuroblastoma) | 9 |
| Colo-205 epithelial cells | C. Henery, Amnis Corp. |
| C3A liver cells | 29 |
| DLS-1 | 13 |
| Embryonic lung fibroblasts | 11 |
| Embyonic neural progenitor cells - undifferentiated and differentiated (neuroepithelial, neuroblastic and glial) | 9 |
| Embryonic stem cells (hES) | 30 |
| FLC4 | 15 |
| HEK 293 | 2,5,15,27 |
| HeLa | 4,5,9,15,18,19 |
| HepG2 (ATCC HB-8065) - hepatocellular carcinoma | 1,2,15,34 |
| Huh7 | 1,4,5,15,19 |
| HUVEC | 28 |
| IMR-32 neuroblastoma (ATCC CCL-127) following differentiation | 4,32 |
| KATO-III (HTB-103) - gastric carcinoma | 4 |
| Keratinocytes | 5 |
| LNCaP - human prostatic adenocarcinoma | F.Matthieu, U. Science et technologies de Lille |
| MCF7 (ATCC HTB-22D) - breast cancer cell line | NIH-NCI |
| MG63 | 5 |
| MRC5 | 7 |
| Mesenchymal stem cells (MSC) (from umbilical chord blood and bone marrow) | 20 |
| CRL-1973 (NTERA-2, Nt-2; malignant pluripotent embryonal carcinoma) | 4, Molecular Probes |
| Pancreatic b-cells | 8 |
| Prenatal cardiomyocytes (hCM) | 18 |
| Primary arterial smooth muscle cells | Molecular Probes |
| Primary chondrocytes | NIH |
| Primary coronary artery endothelia cells (hCEC) | 18 |
| Primary dendritic cells | 25 |
| Primary fibroblasts (hFB) | 18 |
| Primary foreskin fibroblasts (HFF) | 12,28 |
| Primary hepatic stellate cells | 19,23 |
| Primary hepatocytes | 1,2 |
| Primary lung fibroblasts | NIH |
| Primary neural cells | 9 |
| Primary pulmonary arterial endothelial cells (HPAEC) | Molecular Probes |
| Primary umbilical vein endothelial cell (HUVEC) | 28, Molecular Probes |
| Saos-2 | 5,6,19 |
| SHSY-5Y (neuroblastoma) | 27,29 |
| SK-BR-3 | Molecular Probes |
| SK-N-MC | 5 |
| U-2 OS | 27, Molecular Probes |
| U87MG (ATCC HTB-14) - glioma cell line | Genentech |
| W12 | 5 |
| WI38 | 5 |
| Non-human primate cells | |
| COS-7 (African green monkey kidney fibroblast-like cell line) | 3,4,5,9, Molecular Probes |
| CV-1 | 5,15, Molecular Probes |
| Vero | 14, A.Snyder, OHSU |
|
| Cell Types Transduced using BacMam Technology | Reference / Lab |
|---|---|
| Rodents | |
| Hamster cells | |
| CHO (CHO K1, CHO M1WT3, CHO-hIR) | 5,15,31, Molecular Probes |
| Mouse cells | |
| GnRH neuronal cells | S.Singh, Johns Hopkins |
| L929 | 14 |
| Mouse pancreatic b-cells | 8 |
| Mouse primary kidney cells | 22 |
| N2a | 9 |
| NIH 3T3 | Molecular Probes |
| Potoroo (Rat Kangaroo) cells | |
| Ptk2 | M.Davidson, Molecular Expressions Inc. |
| Rabbit cells | |
| CRL-2560 (RH/K30, MT-2; rabbit T-cell line) | 4 |
| Primary hepatocytes | 16 |
| Primary invertebral disc nucleus pulposus cells (in vitro and in vivo) | 24 |
| Rat cells | |
| BHK | 5,15,34 |
| Brain pericytes cell line | Molecular Probes |
| C17.2 cells (differentiated) - multipotent neural stem cell line | 31 |
| Neural stem cells | A.Moutri, The Salk Institute |
| PC12 | 4,5 |
| Primary cerebellar granule neurons | 31 |
| Primary chondrocytes | 21 |
| Primary hepatic stellate cells | 23 |
| Primary hepatocyts | 2 |
| Primary spiral ganglion neurons | 33 |
| Rat2 | M.Davidson, Molecular Expressions Inc. |
| RGM I | 4 |
|
| Cell Types Transduced using BacMam Technology | Reference / Lab |
|---|---|
| Ruminants | |
| Bovine cells | |
| MDB | 5 |
| BT | 5 |
| Ovine (Sheep) cells | |
| FLL-YFT | 5 |
| Cervidae (Deer) cells | |
| Indian Muntjac | M.Davidson, Molecular Expressions Inc. |
| Suidae (Pig) | |
| Suis (Pig) cells | |
| CPK (porcine kindney) | 4,5 |
| FS-L3 (porcine kidney epithelial cell line) | 4,5 |
| PK-15 | 5,34 |
| Left atrial appendage progenitor cells–adult stem cells | M.Rutten and K.Gregory, OMLC |
| Porcine coronary artery smooth muscle cells (pCSMC) | 18 |
| LLC-PK1 | 27, M.Davidson, Molecular Expressions Inc.. |
| Primary Cardiac Smooth Muscle Cells | M.Rutten and K.Gregory, OMLC |
| Carnivores | |
| FoLu (fox lung) | M. Davidson, Molecular Expressions Inc. |
| MDCK (NBL-2; dog kidney) | Molecular Probes |
|
| Cell Types Transduced using BacMam Technology | Reference / Lab |
|---|---|
| Birds | |
| Chicken primary myoblasts | 27 |
| Chicken whole embryonic fibroblast cells | 27 |
| OMK | Molecular Probes |
| Fish | |
| EPC | 26 |
| CHH-1 | 26 |
| Amphibians | |
| Xenopus S3 cell line stably expressing GFP-tubulin | Harvard Univ. |
|
|
We've made it easier
-
Organelle Lights™ delivery is mediated by an insect virus (baculovirus) that is non-infectious to mammalian cells and safe to handle, but still provides a robust cellular delivery. The genetically encoded and prepackaged Organelle Lights™ reagents are ready for immediate use – no need to purify plasmids or worry about vector integrity and quality:
- No lipids, dye-loading chemicals or other potentially harmful treatments are required.
- All Organelle Lights™ batches are pre-packaged at the same concentration, so you don’t have to prepare transfection complexes for each experiment.
- Just add the Organelle Lights™ reagent to your cells for two to four hours, treat with an enhancer, wash, incubate overnight and visualize your results.
Transduction is efficient and reproducible in most cell lines, including primary and stem cells, without apparent cytopathic effects (Table 2). Fluorescent protein expression can therefore be easily titrated to a desired level.
Rugged, persistent, and dependable
The efficient delivery and the genetic content of the Organelle Lights™ marker permits you to transduce a large quantity of primary cells or cell lines in batch mode, aliquot, store frozen, and use as needed, approximating the consistency of stable cell lines without the risk of genetic drift. Fluorescent protein expression is unaffected by the frozen storage. Upon plating transduced cells can remain brightly stained for more than 120 hours.
Specific and precise staining, in combination with the brightness and functional independence of Organelle Lights™ markers, make them ideal for live cell applications—imaging as well as HCS-based. Due to their cellular persistence Organelle Lights™ proteins are easy to multiplex with other fluorescent proteins, organic fluorescent dyes, or QDot® conjugates. Organelle Lights™ proteins are also compatible with fixation and subsequent immunocytochemical processing, such as using anti-GFP antibodies.
Light up cells with powerful and biologically compatible Organelle Lights™ reagents
If your work involves visualizing, tracking, and quantifying dynamic changes in molecules and events in living cells with high spatial and temporal resolution, Organelle Lights™ fluorescent proteins are ideal. The simplicity, ease of use, and reproducibility of Organelle Lights™ proteins lets you concentrate on experimental design, imaging, and data analysis.
|
 | Porcine primary skeletal muscle cell transduced with Organelle Lights™ ER-GFP and Nuc-GFP. Transduction was performed in suspension for 30 minutes and cells were coloaded with Qtracker® 655. Image courtesy of: Ann Bazar, Michael Rutten, and Kenton Gregory, Oregon Medical Laser Center, Bioimaging Suite, Providence St. Vincent Hospital, Portland, Oregon. |
 | U2OS cells transduced with Organelle Lights™ ER-OFP. Cells were imaged in live cell format using a 40X objective. |
 | OK cell transduced with Organelle Lights™ Golgi-GFP. Cells were fixed prior to imaging. Image courtesy of: Michael Davidson, Molecular Expressions Inc., Tallahassee, FL. |
 | FoLu (Gray Fox Lung) cells transduced with Organelle Lights™ Golgi-GFP. Imaging was performed on fixed cells using fluorescence and DIC microscopy. Image courtesy of: Michael Davidson, Molecular Expressions Inc., Tallahassee, FL. |
 | HeLa cells transduced with Organelle Lights™ Mito-GFP and Mito-OFP (pseudocolored red). Cells were counterstained with Hoechst and imaged in live cell format using a 40X objective. |
|
Organelle Lights™ Fluorescent Proteins
-
The stability and brightness of fluorescent proteins, with the efficiency of BacMam delivery—simple, safe, and efficient
|
Table 2 - References for peptide fusion tags
| Organelle Lights™ | Tag description and size | Reference |
|---|---|---|
| Cytoplasm | Nuclear export sequence (1.8 kD) directs cytoplasmic localization | Chevalier, SA (2005) BMC-R 2:1-11 |
| Endoplasmic Reticulum | ER-specific staining is obtained through retention signals from calreticulin & KDEL (1.8 kD N-term & 0.5 kD C-terml) | Fliegel, L (1989) JBC 264:21522-8 |
| Endosomes | Fusion of Emerald GFP to Rab5a (23.8 kD), an early endosomal marker, allowing tracking of endosomal movement within the living cell | Roberts RL (1999) J. Cell Sci. 112:3667-75 |
| Golgi | Human Golgi-resident enzyme N-acetylgalactosaminyl-transferase-2 (12.6 kD) for Golgi-specific live cell labeling | Storrie, B (1998) J.Cell Biol. 143:1505-21 |
| Lysosomes | Fusion of monomeric and bright TagRFP to Lamp1 (lysosomal associated membrane protein; 45 kD) enables tracking of lysosome-mediated process: phagocytosis, endocytosis and autophagy | Falcón-Pérez JM (2005) J Cell Sci. 118:5243-55 |
| Mitochondria | Leader sequence of E1 alpha-pyruvate dehydrogenase (3.1 kD) localizes constructs to the mitochondrial matrix independent of membrane potential | Hanson, G (2004) JBC 279:13044-53 |
| Nuclear Envelope | Nesprin 1 alpha C-terminal transmembrane domain (6.7 kD) targets E-GFP to the nuclear envelope | Zhang, Q (2001) J.Cell Sci. 114:4485-98 |
| Nucleus | SV40 nuclear localization sequence (1.0 kD) restricts fluorescent protein expression to the nucleus | Dingwall, C (1991) TiBS 16:478-81 |
| Peroxisome | Peroxisomal C-terminal targeting sequence (0.3 kD) enables staining and tracking of peroxisomes in live cells | Gould, SJ (1989) J.Cell Biol. 108:1657-64 |
| Plasma Membrane | Myristoylation/palmitoylation sequence from Lck tyrosine kinase (0.9 kD) mediates staining of the plasma membrane | Kabouridis, PS (1997) EMBO J. 16:4983-98 |
| Synaptophysin | Synaptic vesicle protein involved in vesicle endo- and exocytosis (33.8 kD, N terminus of GFP) | Kaether, C (2000) MBC 11:1213-1224 |
| Null virus (control) | Cellular Lights™ Null virus lacks mammalian genetic elements and can be used as a control to determine potential baculovirus-mediated effects | N/A |
|
Table 3 - Typical transduction efficiencies obtained by using standard Organelle Lights™ protocol
| Cell line | Transduction efficiency |
|---|---|
| CHO, CHO K1, CHO M1WT3, etc. | 70% - 80% |
| COS-7 | >90% |
| CRL-1973 (NTERA-2) | 30% |
| HEK 293 | >90% |
| HeLa | 60% - 70% |
| HepG2 | >90% |
| Indian Muntjac | 80% - 90% |
| Left atrial appendage progenitor cells–adult porcine stem cells | 40% - 50% |
| NIH 3T3 | 40% - 50% |
| OMK | 50% - 60% |
| Primary Cardiac Smooth Muscle Cells | 40% - 50% |
| SK-BR-3 | 40% - 50% |
| U-2 OS | >90% |
| WI38 | 30% - 40% |
| Cells of hematopoiteic lineages | Not readily amenable |
|
Table 4
| References | |
|---|---|
| 1 | Hofmann, C. et al. (1995) Proc. Natl. Acad. Sci. USA 92:10099-10103 |
| 2 | Bouce, F.M. & Boucher, N. (1996) Proc. Natl. Acad. Sci. USA 93:2348-2352 |
| 3 | Yap, C.-C. et al. (1997) Virology 231:192-200 |
| 4 | Shoji, I. et al. (1997) J. Gen. Virol. 78:2657-2664 |
| 5 | Condreay, J.P. et al. (1999) Proc. Natl. Acad. Sci. USA 96:127-132 |
| 6 | Merrihew, R.V. et al. (2001) J. Virol. 75:903-909 |
| 7 | Palambo, F. et al. (1998) J. Virol. 72:5025-5034 |
| 8 | Ma, L. et al. (2000) Diabetes 49:1986-1991 |
| 9 | Sarkis, C. et al. (2000) Proc. Natl. Acad. Sci USA 98:14638-14643 |
| 10 | Ye, G.J. et al. (2000) J. Virol. 74:1355-1363 |
| 11 | Lopez, P. et al. (2001) J. Virol. 75:3832-3840 |
| 12 | Dwarakanath, R.S. et al. (2001) Virology 284:297-307 |
| 13 | Barsoum, J. et al. (1997) Hum. Gene Ther. 8:2011-2018 |
| 14 | Airenne, K.J. et al. (2000) Gene Ther. 7:1499-1504 |
| 15 | Tani, H. et al. (2001) Virology 279:343-353 |
| 16 | Munger, J. & Roizman, B. (2001) Proc. Natl. Acad. Sci. USA 98:10410-10415 |
| 17 | Ames, R.S. et al. (2004) Receptors Channels 10:117-124 |
| 18 | Grassi, G. et al. (2006) Arch. Virol. 151:255-271 |
| 19 | Nicholson, L. et al. (2004) Mol. Ther. 11:638-644 |
| 20 | Ho, Y.-C. et al. (2005) J. Gene Med. 7:860-868 |
| 21 | Ho, Y.-C. et al. (2004) Biotechnol. Bioeng. 88:643-651 |
| 22 | Liang, C.Y. et al. (2004) Arch. Virol. 149:51-60 |
| 23 | Gao, R. et al. (2002) Liver 22:15-22 |
| 24 | Liu, X. et al. (2006) Spine 31:732-5 |
| 25 | Strauss, R. et al. (2007) Mol. Ther. 15:193-202 |
| 26 | Leisy, D.J. et al. (2003) J. Gen. Virol. 84:1173-8 |
| 27 | Hassan, N.J. (2006) Prot. Expr. Purif. 47:591-8 |
| 28 | Kronschnabl, M. & T. Stamminger (2003) J. Gen. Viro. 84:61-73 |
| 29 | Andersson, M. et al. (2007) BMC Cell Biol. 8:6 |
| 30 | Zheng, J. et al. (2007) Stem Cells 25:1055 |
| 31 | Li, Y. et al. (2004) Exp. Physiol. 90:39-44 |
| 32 | Näsman, J. et al. (2006) J. Neurosci. 26:10658-10666 |
| 33 | Wang, J. et al. (2006) NeuroReport 18:1329-1333 |
| 34 | Gao, H. et al. (2007) J. Biotechnol. 131:138-143 |
|
|
Organelle Lights™ ER-GFP
Cos cells transduced with Organelle Lights™ ER-GFP.
