FAP-Tagging Kit for Live Cell Assays (manual)
Vector Sequences and Annotation: pMFAPalpha2; pMFAPbeta1
SpectraGenetics offers flexible, easy to use vectors for tagging cells and proteins with a series of new fluorescent reporters called fluorogen-activating proteins (FAPs)1.
- FAP-tags® exhibit fluorescence only in the presence of submicromolar concentrations of cognate small-molecule fluorogens. The fluorescent FAP/fluorogen complex is called a fluoromodule. Neither the FAP nor the fluorogen is fluorescent by itself.
- FAP-tags® allow the user to turn the fluorescent signal on and off by adding or removing fluorogen, or to change the signal wavelength by substituting one fluorogen for another.
Spectragenetics has developed two different types of FAPs, α and β. Experiments can be designed with both FAPs to measure receptors internalization simultaneously (multiplex) or an individual FAP can be used for more detailed trafficking experiments. Each FAP has a unique set of available fluorogens. The α FAP has cell impermeant, permeant, pH sensitive and extra bright available. This allows measurements of protein at the surface, total protein, internalization 1 color), and trafficking to and from the cell membrane. The βFAP has 2 spectrally distinct dyes that allow for measurement of surface protein, internalization (2 color), re-sensitization and pulse/chase experiments. Fluoromodule Properties:
| FAP | Fluorogen | Excitation Max | Emission Max | Kd | Recommended fluorogen working concentration |
| FAPα2 | αRED1-np1 | 631 nm | 650 nm | < 1 nM | 100 nM |
| αRED1-p1* | 640 nm | 668 nm | < 1 nM | 100 nM | |
| FAPβ1 | βGREEN-np1 | 509 nm | 530 nm | 3.1 nM | 100 nM |
| βRED-np1 | 509,650 nm | 670 nm | ~13nM | 100nM |
* Membrane permeant fluorogen. All other fluorogens are membrane-impermeant. When used in conjunction with membrane-impermeable fluorogens, FAP-tags® have proved particularly useful in visualizing the plasma membranes of cultured cells2, and in live-cell assays that monitor the translocation of membrane proteins to or from the cell surface3,4. SpectraGenetics offers vectors for FAP-tagging the plasma membrane of mammalian cells and cloning kits for expressing FAP-tagged fusion proteins, as well as an expanding collection of transfection-ready FAP-tagged genes. FAP-Tagging kits for mammalian surface expression of FAPs. The vectors in these kits express FAPs that localize to the plasma membrane of transfected cells. The vectors can also be used to clone and express cell-surface or secreted proteins with extracellular N-terminal FAP tags.
|
FAP-tag® In vector |
Product |
Price |
| FAPα2 | pMFAPα2-kit-001 | $499 |
| FAPβ1 | pMFAPβ1-kit-001 | $499 |
Each kit contains a supply of fluorogen sufficient for 20 experiments with a media volume of 1 ml each, or 100 experiments with a volume of 200 µL each.
Fluorogen Pack Sizes and Prices:
α |
Fluorogen | Catalog | Amount (nmol) | Measurements | Price |
| αRED-np | αRED-np-001 | 2 | 100 | $199 | |
| αRED-np-010 | 20 | 1,000 | $999 | ||
| αRED-np-100 | 200 | 10,000 | $4999 | ||
| αRED-p | αRED-p-001 | 2 | 100 | $199 | |
| αRED-p-010 | 20 | 1,000 | $999 | ||
| αRED-p-100 | 200 | 10,000 | $4999 | ||
| αRED-Bright | se-Dyedron5x-001 | 2 | 100 | $399 | |
| se-Dyedron5x-010 | 20 | 1,000 | $1999 | ||
| se-Dyedron5x-100 | 200 | 10,000 | $9999 | ||
| αRED-pH | se-RED-pH-001 | 2 | 100 | $399 | |
| se-RED-pH-010 | 20 | 1,000 | $1999 | ||
| se-RED-pH-100 | 200 | 10,000 | $9999 | ||
β |
Fluorogen | Catalog | Amount (nmol) | Measurements | Price |
| βGREEN-np | βGREEN-np-001 | 2 | 100 | $199 | |
| βGREEN-np-010 | 20 | 1,000 | $999 | ||
| βGREEN-np-100 | 200 | 10,000 | $4999 | ||
| βRED-np | βRED-np-001 | 2 | 100 | $299 | |
| βRED-np-010 | 20 | 1,000 | $1499 | ||
| βRED-np-100 | 200 | 10,000 | $7499 |
α fluorogens are used with αFAPs whereas β fluorogens are used with β FAPs.