| Bioactivity | Flubida-2 is a cell permeable dye which can be hydrolyzed to Fubi-2 by endoesterases in cells (after hydrolysis, Ex=492 nm, Em=517 nm). Flubida-2 can be used to detect pH at a specific site in a cell organelle by directing the probe to where avidin fusion proteins are located[1]. |
| Invitro | Guidelines (Following is our recommended protocol. This protocol only provides a guideline, and should be modified according to your specific needs)[1].1. Dissolve Flubida-2 in DMSO (approximately 2 mM). 2. Mix the stock solution 1: 1 with 20% (w/v in DMSO) Pluronic F-127 (Molecular Probes), and dilute to the desired final concentration (2-4 μM) with serum-free DMEM. 3. 30 to 48 h post-transfection with either AV-KDEL or ST-AV DNA, HeLa cells are rinsed once with serum-free DMEM and loaded with 2-4 μM Flubida-2 for 3-5 h (or overnight for 10-15 h). 4. Chase the labeled cells with normal growth medium for at least 2 h, to allow excess dye-biotin to exit from the cytosol. 5. The strong avidin-biotin interaction ensures stable, specific avidin-Flubi- binding that resisted washing. Biotin starvation of the cells is not necessary before Flubida- loading, as staining was bright and stable. |
| Name | Flubida-2 |
| Formula | C86H96N8O20S2 |
| Molar Mass | 812.93 |
| Transport | Room temperature in continental US; may vary elsewhere. |
| Storage | Please store the product under the recommended conditions in the Certificate of Analysis. |
| Reference | [1]. M M Wu, et al. Studying organelle physiology with fusion protein-targeted avidin and fluorescent biotin conjugates. Methods Enzymol. 2000;327:546-64. |