Cell Navigator® Live Cell Tubulin Staining Kit
Ordering information
| Price | |
| Catalog Number | |
| Unit Size | |
| Quantity |
Additional ordering information
| Telephone | 1-800-990-8053 |
| Fax | 1-800-609-2943 |
| sales@aatbio.com | |
| Quotation | Request |
| International | See distributors |
| Shipping | Standard overnight for United States, inquire for international |
Storage, safety and handling
| Certificate of Origin | Download PDF |
| Intended use | Research Use Only (RUO) |
Related products
| Overview |  SDSProtocol |
See also: Cell Structures and Organelles, Mitochondria
Cell Navigator® Live Cell Tubulin Staining Kit provides a robust method to fluorescently image tubulins in live cells with Tubulite™ Red. The probe is permeant to live cells, thus does not require cells to be fixated for imaging tubulins. It can be conveniently used for tracking tubulin polymerization process in live cells. Its red spectral wavelengths and good cell permeability make this probe readily to use with other colors such as GFP expressed cells or nuclear dyes such as DAPI. The neutral Tubulite™ Red readily passes through the plasma membranes of live cells. Once inside the cells, the lipophilic blocking group is cleaved by esterases, resulting into a negatively charged product, which is well retained inside the cells.
Platform
Fluorescence microscope
| Excitation | Cy5 filter set |
| Emission | Cy5 filter set |
| Recommended plate | Black wall/clear bottom |
Components
Example protocol
AT A GLANCE
Protocol Summary
- Prepare cells with test compounds at a density of 5 × 105 to 1 × 106 cells/mL
- Prepare and add TubuliteTM Red working solution to cells
- Incubate at 37 °C for 30 to 60 minutes
- Read fluorescence intensity with Cy5 filter set
Tubulite™ Red does not stain formaldehyde fixed cells. Cells can not be fixed after staining with Tubulite™ Red as fixation alters the structure of micotubules.
CELL PREPARATION
For guidelines on cell sample preparation, please visit https://www.aatbio.com/resources/guides/cell-sample-preparation.html
PREPARATION OF STOCK SOLUTIONS
Unless otherwise noted, all unused stock solutions should be divided into single-use aliquots and stored at -20 °C after preparation. Avoid repeated freeze-thaw cycles.
TubuliteTM Red stock solution (500X)
Add 25 µL DMSO (Component D) into the vial of TubuliteTM Red (Component A), and mix well. Note: Aliquot and stored the unused TubuliteTM Red stock solution at -20 °C. Avoid repeated freeze/thaw cycles.PREPARATION OF WORKING SOLUTION
TubuliteTM Red working solution (1X)
Add 2.5 µL of TubuliteTM Red stock solution stock solution and 100 µL 25 mM ReadiUse™ probenecid (Component D) into 1 mL of Assay Buffer (Component B) or buffer of your choice, and mix well. Note: We recommend making TubuliteTM Red working solution fresh for every use. The working solution is stable for several hours.SAMPLE EXPERIMENTAL PROTOCOL
- Prepare cell samples as per need.
- Remove the cell growth medium and wash cells with PBS (Not provided) or any other buffer of your choice. (Optional).
- Add 100 µL TubuliteTM Red working solution and incubate them at 37 °C incubator for 30 to 60 minutes. Note: The appropriate incubation time depends on the individual cell type and cell concentration used. Optimize the incubation time for each experiment.
- Remove the working solution and wash cells twice with PBS or any other buffer of your choice with 2.5 mM probenecid (diluted from Component C).
- Cover cells with Assay Buffer with 2.5 mM probenecid (diluted from Component C) and monitor the fluorescence intensity with fluorescence microscope using Cy5 filter set.
Images
Figure 1. Imaging of tubulins in live HeLa cells. HeLa cells were co-labelled with TubuliteTM Red and DAPI (Cat# 17507) for 60 minutes in a 37 oC, 5% CO2 incubator. The fluorescence image was taken with a fluorescence microscope (Cy5 filter set).