For Your Copy of the New 2012 Cytoskeleton Product Catalogue
Please contact us!
About Acti-stain™
The Acti-stain™ line of fluorescent phalloidins have been developed with an emphasis on creating exceptionally bright and stable probes at an economical price. Side-by-side comparisons with leading competing products demonstrates that you will enjoy savings while not sacrificing the quality of the stain when switching to an Acti-stain™ probe. Additionally these probes are compatible with popular filter sets such as FITC, TRITC and Cy5. The combination of in-house manufacturing, stringent quality control and convenient packaging provides a great value. Give us a try and see for yourself.
Product Uses Include
- Stain F-actin in fixed cells
- Stabilize actin filaments in vitro
- Visualize actin filaments in vitro
Actin staining is very useful in determining the structure and function of the cytoskeleton in living and fixed cells. The actin cytoskeleton is a very dynamic and labile structure in the living cell, but it can be fixed with paraformaldehyde prior to probing or staining for actin structures.
| Acti-stain™ 488 phalloidin Cat. # PHDG1 
|
Acti-stain™ 555 phalloidin Cat. # PHDH1 
|
Acti-stain™ 670 phalloidin Cat. # PHDN1 
|
|||
| Swiss 3T3 cell stained with anti-vinculin (red), Dapi (blue nucleus) and F-actin is stained with Acti-stain™ 488 (green F-actin, Cat.# PHDG1).
|
Swiss 3T3 cell with activated Rho, nucleus is stained with Dapi (blue) and F-actin is stained with Acti-stain™ 555 (red F-actin, Cat.# PHDH1).
|
Mitotic Swiss 3T3 cell, F-actin stained with Acti-stain™ 670 (far-red, Cat. # PHDN1), nuclear DNA stained with Dapi (blue).
|
|||
| Amount | Price | Amount | Price | Amount | Price |
| 300 Slides* |
£145.80 | 300 Slides* | £145.80 | 300 Slides* |
£145.80 |
| *One slide equals enough phalloidin to stain a 25 mm2 coverslip. | |||||
NOTE OFFER : Buy Any Two Phalloidins and get a Third One Free! (Expires 3rd June 2011)
|
Specifications: |
||
 |
 |
 |
|
Absorbance and fluorescence scan of Acti-stain™ 488. Labeled phalloidin was diluted into methanol and its absorbance and excitation spectra were scanned between 350-750 and 500-750 nm, respectively. Absorbance peaks at 500 nm and fluorescence at 550 nm. |
Absorbance and fluorescence scan of Acti-stain™ 555. Labeled phalloidin was diluted into methanol and its absorbance and excitation spectra were scanned between 300-750 and 560-750 nm, respectively. Absorbance peaks at 560 nm and fluorescence at 575 nm. |
Absorbance and fluorescence scan of Acti-stain™ 670. Labeled phalloidin was diluted into methanol and its absorbance and excitation spectra were scanned between 300-750 and 600-750 nm, respectively. Absorbance peaks at 625 nm and fluorescence at 675 nm. |
Application Notes:
Actin Staining in Fixed Cells
In fixed cells, actin structures can be visualized by actin antibodies (1), fluorescent phalloidins (2) such as Cytoskeleton’s Acti-stain™ product line, or electron microscopy (3). Fluorescent phalloidins only binds to the native quaternary structure of F-actin, therefore paraformaldehyde must be used as the fixative because it retains protein structure. Methanol destroys native conformation and hence is not suitable for actin staining with phalloidin.
Fixed cell actin staining products:
Acti-stain™ 488 (green fluorescence fixed cell stain)
Acti-stain™ 555 (red fluorescence fixed cell stain)
Acti-stain™ 670 (far-red fluorescence fixed cell stain)
Rhodamine Phalloidin (red fluorescence fixed cell stain)
F-actin Visualization Biochem Kit™ (rhodamine)
Actin antibody (rabbit polyclonal)
Acti-stain™ 488
(green fluorescence fixed cell stain)
Swiss 3T3 cells were stained with Acti-stain™488. F-actin filaments are clearly seen at the periphery of the cells and as stress fibers in the cytoplasm.
Actin Staining in Living Cells
In living cells actin structures can be investigated by introduction of a fluorescent actin fusion protein or a fluorescently labeled actin binding protein sub-domain. Fluorescent actin is the most accurate reporter of actin structures but it is sometimes difficult to introduce these reporters into cells. Traditionally this is accomplished by microinjection or protein transfection. Additionally, F-actin binding protein tags are useful conjugates to probe F-actin in live cells (5,6). While some cells are easily transfected, others, e.g. primary cells, are currently difficult to transfect. This will undoubtedly improve over time with improved methods to introduce proteins and/or DNA constructs.
Living cell actin staining products:
Rhodamine actin (muscle actin)
Rhodamine actin (non-muscle actin)Rhodamine Actin
(Living Cell Actin Stain)
Rhodamine actin has been microinjected into CHO cells. The labeled actin rapidly incorporates into the cellular actin cytoskeleton and allows real time observation of actin dynamics.
If you have any questions about actin staining, please contact Cytoskeleton Technical Support.
References:
1. Lazarides E, and Weber K. 1974. Actin antibody: The specific visualization of actin filaments in non-muscle cells. PNAS USA 71, 2268-2272.
2. Wulf F, Deboben A, Bautz FA, Fualstich H, and Wieland TH. 1979. Fluorescent phallotoxin, a tool for the visualization of cellular actin. PNAS USA71, 4498-4502.
3. Weber K, Rathke PC, & Osborn M. (1978). Cytoplasmic microtubular images in glutaraldehyde-fixed tissue culture cells by electron microscopy and by immunofluorescence microscopy. PNAS USA75, 1820-1824.
4. Riedl J, Crevenna AH, Kessenbrock K, Haochen Yu J, Neukirchen D, Bista M, Bradke F, Jenne D, Holak TA, Werb Z, Sixt M. and Wedlich-Soldner R. 2008. Lifeact: a versatile marker to visualize F-actin. Nature Methods, 5(7), 605-611.