Product Uses Include
Phalloidin's usefulness as a laboratory tool is well established and lies in it's ability to inhibit microfilament de-polymerization. Thus, phalloidin stabilized microfilaments are used as substrates for the identification and characterization of the ever increasing number of microfilament associated proteins. In addition, rhodamine phalloidin can be used for staining the actin cytoskeleton in cells, and also as a fluorescent marker/stabilizer for microfilaments used in motility assays.
The rhodamine phalloidin is provided as a lyophilized powder. When reconstituted with 500 µl methanol, it is at 14 µM (200 x stock).
In keeping with our policy of providing only the highest quality products, we are now pleased to inform you that Cytoskeleton is offering rhodamine phalloidin at a chromatographic purity of >99%. This is the purest phalloidin commercially available for research use.
We have determined that the microfilament stabilizing property of our product is equal or superior to any other commercially available phalloidins. Microfilaments in 70 nM phalloidin are stable at room temperature for over one week.
Figure 1. A Swiss 3T3 fibroblast stained with rhodamine phalloidin (PHDR1). The phalloidin binds specifically to F-actin and does therefore stain the actin stress fibers in the cell
Lin, L., et al. Phase separation-mediated condensation of Whirlin-Myo15-Eps8 stereocilia tip complex https://doi.org/10.1016/j.celrep.2021.108770 (2021)
Danielsson, B. E. et al. Lamin microaggregates lead to altered mechanotransmission in progerin-expressing cells. Nucleus 11, 194–204 (2020).
Yang, S. et al. Self-assembling peptide hydrogels functionalized with LN- And BDNF- mimicking epitopes synergistically enhance peripheral nerve regeneration. Theranostics 10, 8227–8249 (2020).
Zhu, J. et al. Bioactive poly (methyl methacrylate) bone cement for the treatment of osteoporotic vertebral compression fractures. Theranostics 10, 6544–6560 (2020).
Lu, F., Sun, X., Xu, X. & Jiang, X. SILAC-based proteomic profiling of the suppression of TGF-β1-induced lung fibroblast-to-myofibroblast differentiation by trehalose. Toxicol. Appl. Pharmacol. 391, 114916 (2020).
Huang, Yuxing et al. “Arp2/3-Branched Actin Maintains an Active Pool of GTP-RhoA and Controls RhoA Abundance.” Cells vol. 8,10 1264. 16 Oct. 2019, doi:10.3390/cells8101264
Shan, Lijun et al. “In Situ Controlled Surface Microstructure of 3D Printed Ti Alloy to Promote Its Osteointegration.” Materials (Basel, Switzerland) vol. 12,5 815. 10 Mar. 2019, doi:10.3390/ma12050815
Zhang, Ying et al. “Effects of testosterone on the expression levels of AMH, VEGF and HIF-1α in mouse granulosa cells.” Experimental and therapeutic medicine vol. 12,2 (2016): 883-888. doi:10.3892/etm.2016.3436
Lee et al., 2013. Phellinus baumii extract influences pathogenesis of Brucella abortus in phagocyte by disrupting the phagocytic and intracellular trafficking pathway. J. Appl. Microbiol. 114, 329–338.
Baruzzi and Berton, 2012. The tyrosine kinase Abl is a component of macrophage podosomes and is required for podosome formation and function. E. J. Immunol. doi: 10.1002/eji.201142270.
Shiozaki et al., 2012. XB130 as an Independent Prognostic Factor in Human Esophageal Squamous Cell Carcinoma. Ann. Surg. Oncology. doi: 10.1245/s10434-012-2474-4.
Gallo et al., 2012. Regulation of the Actin Cytoskeleton by Rho Kinase Controls Antigen Presentation by CD1d. J. Immunol. doi: 10.4049/jimmunol.1101484.
Lee et al., 2012. Dysadherin expression promotes the motility and survival of human breast cancer cells by AKT activation. Cancer Sci. v 103, pp 1280-1289.
Shi et al., 2012. MTA-enriched nanocomposite TiO2-polymeric powder coatings support human mesenchymal cell attachment and growth. Biomed. Mater. 7:055006. doi:10.1088/1748-6041/7/5/055006.
Ezratty et al., 2005. Microtubule-induced focal adhesion disassembly is mediated by dynamin and focal adhesion kinase. Nat. Cell Biol. v 7, pp 581-590.
Gomes et al., 2005. Nuclear movement regulated by Cdc42, MRCK, myosin, and actin flow establishes MTOC polarization in migrating cells. Cell. v 121, pp 451-463.
Chandhoke et al., 2004. β3 integrin phosphorylation is essential for Arp3 organization into leukocyte αVβ3-vitronectin adhesion contacts. J. Cell Sci. v 117, pp 1431-1441.
Hsieh-Wilson et al., 2003. Phosphorylation of spinophilin modulates its interaction with actin filaments. J. Biol. Chem. v 278, pp 1186-1194.
Qian et al., 2002. PKC phosphorylation increases the ability of AFAP-110 to cross-link actin filaments. Mol. Biol. Cell. v 13, pp 2311-2322.
Chen, J., Fabry, B., Schiffrin, E. L. and Wang, N. (2001). Twisting integrin receptors increases endothelin-1 gene expression in endothelial cells. Am. J. Physiol. Cell Physiol. v 280, pp C1475-C1484.
Question 1: Can I use fluorescently-labeled phalloidin to stain F-actin in living cells?
Answer 1: Unfortunately, no, phalloidin cannot be used to stain F-actin in living cells. Phalloidins are used to stain F-actin in fixed cells. Fluorescent phalloidins only bind to the native quaternary structure of F-actin which provides a low background. The correct fixation condition for phalloidin binding is 3.7% (v/v) paraformaldehyde in PBS for 10 min because it retains the quaternary protein structure which is necessary for high affinity binding of phalloidin. Methanol fixation destroys the native conformation and hence is not suitable for F-actin staining with phalloidin. To monitor actin dynamics in living cells, micro-injection of rhodamine-labeled actin (Cat. # APHR or AR05) is recommended. Please see those datasheets for more information.
Question 2: Which of the fluorescently-labeled phalloidin is the most stable/brightest?
Answer 2: The brightest and most stable of the Acti-stains is Acti-stain 488 (green fluorescence; Cat. # PHDG1). Please see the table below for additional information on all of our Acti-stains.
Stability to photobleaching* (1/2 life in sec)
Background (% of total AFU at 100 nM)
* = measured in the absence of antifade.
If you have any questions concerning this product, please contact our Technical Service department at firstname.lastname@example.org.