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Organelle Marker FAQs
1. Q: What are the fluorescent proteins are used for the organelle markers?
A: The fluorescent proteins are turboGFP and turboRFP from Evrogen. Both proteins are super bright with brightness % compared to EGFP of 112 and 187, respectively. They are both fast maturing, allowing detection of fluorescence as early as 8-10 hrs post transfection. In spite of their dimeric structures, tGFP and tRFP have been successfully used as fusion protein for subcellular structure labeling. All OriGeneGÇÖs organelle markers are tested individually for correct labeling
2. Q: What is the excitation/emission wavelength of tGFP?
A: The excitation/ emission max = 482/ 502 nm. tGFP can be detected using common fluorescence filter sets for EGFP, FITC, and other green dyes. Recommended Omega Optical filter sets are QMAX_Green, XF100_2, XF100_3, XF115_2, and XF116_2
3. Q: What is the excitation/emission wavelength of RFP?
A: The excitation/emission max=553/574 nm. tRFP can be detected using common fluorescence filter sets for Texas Red. Recommended Omega Optical filter sets are QMAX_Yellow, XF108_2, XF101_2, and XF111_2. tRFP can also be detected using TRITC filter set.
4. Q: Can I use a regular fluorescent microscope to obtain organelle images?
A: Conventional fluorescent microscope can be used for viewing some of the organelle or subcellular structures. However the images are usually lack the desired clarity. Confocal microscope
5. Q: What antibody can I use to detect the tGFP fusion protein on a Western blot?
6. Q: What antibody can I use to detect the RFP fusion protein on a Western blot?
7. Q: Can I make stable mammalian cell lines with the organelle markers?
A: Yes. All organelle marker plasmids carry a neomycin resistance marker for G418 selection.
8. Q: What positive control should I use with these markers?
A: Optional nuclear protein marker, such as cyclin D1, with additional cost. If you have an antibody specific for an organelle, you can also use that as a reference point.
9. Q: Where can I find the sequence and cloning sites for my Organelle marker?
10. Q: Which vector should I use as a negative control for my GFP-tagged or RFP-tagged marker?
11. Q: Can I shuttle my ORF using your PrecisionShuttle system to change my fluorescent tag?
12. Q: How are the organelle marker tested?
13. Q: I am writing a paper for publication and need to describe this product. How should I cite?
A: We recommend that you refer to the product by its specific catalog number and refer to us as OriGene Technologies (Rockville, MD). Furthermore, we'd love to hear from you when your paper is published. Inform us and we will send a gift.