For research use only. Not for therapeutic Use.
5(6)-TAMRA is a fluorescent dye molecule widely used as a label for peptides and proteins.
1. Peptide Labeling:
The polypeptide R9 was incubated with TAMRA (4-equivalent) in dimethylformamide containing benzotriazole-1-acyloxy-tripyrodiene-phosphohexafluorophosphate (4-equivalent) and diisopropylethylamine (8-equivalent) for 4 hours and labeled on the resin. Wash the resin with dimethylformamide and methylene chloride, and then de-protect as described above.
2. Peptide Internalization:
CHO cells were inoculated with Ham’s F-12 medium in a 4-well Lab-Tek-II chamber slide (Nalge Nunc International, Naperville, IL). Labeled peptides were added to each slide and incubated at 37°C. After 60 minutes, cells were washed with PBS containing Ca2+ (0.1 g/L) and Mg2+ (0.1 g/L) (6 times). Fluorescence microscope observation.
3. Heparin-Affinity Chromatography:
The labeled peptide (~ 2mg) was loaded onto the column and eluted in the same buffer with a linear gradient of NaCl (0-2 M). Peptide elution was monitored at 280 nm and 535 nm absorbance.
4. Binding of TAMRA–R9 to Heparin:
When TAMRA-R9 binds to soluble heparin, fluorescence enhancement at 590 nm (excitation:531 nm).
| Catalog Number | I013841 |
| CAS Number | 98181-63-6 |
| Synonyms | 3′,6′-bis(dimethylamino)-1-oxospiro[2-benzofuran-3,9′-xanthene]-5-carboxylic acid;3′,6′-bis(dimethylamino)-3-oxospiro[2-benzofuran-1,9′-xanthene]-5-carboxylic acid |
| Molecular Formula | C25H22N2O5 |
| Purity | ≥95% |
| InChI | InChI=1S/2C25H22N2O5/c1-26(2)15-6-9-19-21(12-15)31-22-13-16(27(3)4)7-10-20(22)25(19)18-8-5-14(23(28)29)11-17(18)24(30)32-25;1-26(2)15-6-9-18-21(12-15)31-22-13-16(27(3)4)7-10-19(22)25(18)20-11-14(23(28)29)5-8-17(20)24(30)32-25/h2*5-13H,1-4H3,(H,28,29) |
| InChIKey | LBUFYZITRTUEFI-UHFFFAOYSA-N |
| SMILES | CN(C)C1=CC2=C(C=C1)C3(C4=C(C=C(C=C4)C(=O)O)C(=O)O3)C5=C(O2)C=C(C=C5)N(C)C.CN(C)C1=CC2=C(C=C1)C3(C4=C(O2)C=C(C=C4)N(C)C)C5=C(C=CC(=C5)C(=O)O)C(=O)O3 |
| Reference | [1]. Fuchs SM, et al. Pathway for polyarginine entry into mammalian cells. Biochemistry. 2004 Mar 9;43(9):2438-44. [2]. Gerber D, et al. Insertion and organization within membranes of the delta-endotoxin pore-forming domain, helix 4-loop-helix 5, and inhibition of its activity by a mutant helix 4 peptide. J Biol Chem. 2000 Aug 4;275(31):23602-7. [3]. Vinayak R, et al. Automated, solid-phase coupling of rhodamine dye acids to 5′ amino oligonucleotides. Nucleic Acids Symp Ser. 2000;(44):257-8. |
