| 包装: | 1mg |
| 规格: | 98% |
| 市场价: | 7835元 |
| 分子量: | 1231.4 |
Background:
TRITC is an orange-red fluorescent probe，Phalloidin, a cyclic heptapeptide toxin derived from Amanita phalloides, selectively binds filamentous actin F-actin with high affinity (Kd= 20 nM), but not monomeric actin g-actin[1 2]. TRITC Phalloidin staining has strong specificity and high contrast. TRITC Phalloidin is widely used to immobilize permeable cells, but it can also enter living cells..
TRITC Phalloidin staining showed that MCF-7 cells transformed from round to irregular polygon with stiffness increase either in normoxia or hypoxia[5]. Cytoskeletal analysis by TRITC Phalloidin staining show that HeLa cells in the mixed hydrogel beads closely link to each other[6].The cDNAs coding for IEF's 8118(human homolog of bovine GDI ) and 8120(a distinct although related protein) were recombined into vaccinia virus and expressed in differentiated human keratinocytes and their effect on the actin cytoskeleton was assessed by immunofluorescence using TRITC Phalloidin. The results showed that overexpression of both GDI proteins leads to rounding up of the cells and loss of stress fibers and focal contact sites[4].Estrogen is known to have a direct effect on bone forming osteoblasts and bone resorbing osteoclasts. The cells were cultured in either medium alone or medium supplemented with β-estradiol and then subjected to Atomic Force Microscopy indentation (AFM) to determine elastic modulus. The underlying changes in cytoskeleton were studied by staining the cells with TRITC Phalloidin. With estradiol treatment, elastic modulus of osteoblasts significantly decreased by 43-46%[7]
In Amoeba proteus (strain B),When used TRITC Phalloidin staining to examine the presence of F-actin in the amoeba nucleus. No significant amount of F-actin was detected in the nucleus; instead, F-actin formed a cytoplasmic meshwork of filaments and bundles supporting the nucleus[3]
参考文献:
[1]: Bereiter-Hahn J, Kajstura J. Scanning microfluorometric measurement of TRITC-phalloidin labelled F-actin. Dependence of F-actin content on density of normal and transformed cells. Histochemistry. 1988;90(4):271-6. doi: 10.1007/BF00495970. PMID: 3230049.
[2]: Cano ML, Cassimeris L, et,al. Characterization of tetramethylrhodaminyl-phalloidin binding to cellular F-actin. Cell Motil Cytoskeleton. 1992;21(2):147-58. doi: 10.1002/cm.970210208. PMID: 1559266.
[3]: Berdieva M, Bogolyubov D, et,al. Nucleus-associated actin in Amoeba proteus. Eur J Protistol. 2016 Oct;56:191-199. doi: 10.1016/j.ejop.2016.09.002. Epub 2016 Sep 9. PMID: 27684042.
[4]: Leffers H, Nielsen MS, et,al. Identification of two human Rho GDP dissociation inhibitor proteins whose overexpression leads to disruption of the actin cytoskeleton. Exp Cell Res. 1993 Dec;209(2):165-74. doi: 10.1006/excr.1993.1298. PMID: 8262133.
[5]: Lv Y, Chen C, Zhao B, et,al. Regulation of matrix stiffness on the epithelial-mesenchymal transition of breast cancer cells under hypoxia environment. Naturwissenschaften. 2017 Jun;104(5-6):38. doi: 10.1007/s00114-017-1461-9. Epub 2017 Apr 5. PMID: 28382476.
[6]: Wang Y, Wang J. Mixed hydrogel bead-based tumor spheroid formation and anticancer drug testing. Analyst. 2014 May 21;139(10):2449-58. doi: 10.1039/c4an00015c. PMID: 24699505.
[7]: Muthukumaran P, Lim CT, et,al. Estradiol influences the mechanical properties of human fetal osteoblasts through cytoskeletal changes. Biochem Biophys Res Commun. 2012 Jul 6;423(3):503-8. doi: 10.1016/j.bbrc.2012.05.149. Epub 2012 Jun 5. PMID: 22683634.
Protocol:
| Cell experiment [1]: | |
Cell lines | Human breast cancer cells MCF-7 |
Preparation Method | Cells were cultured on polyacrylamide gels with different stiffness under normoxia or hypoxia condition for 48 h, after a series of treatments, The cells were washed with PBS and incubated with TRITC Phalloidin (1:200) for 2 h |
Reaction Conditions | The cells were incubated with TRITC Phalloidin (1:200) for 2 h |
Applications | TRITC-phalloidin staining showed that MCF-7 cells transformed from round to irregular polygon with stiffness increase either in normoxia or hypoxia. |
| Animal experiment [2]: | |
Animal models | Amoeba proteus (strain B) |
Preparation Method | Squash preparations were used for staining. After fixation and permeabilization, washed cells were incubated in 35 μg/ml TRITC Phalloidin solution in a moist chamber at room temperature for 30 min. The preparations were mounted in Vectashield with 1 μg/ml DAPI. |
Dosage form | 35 μg/ml TRITC Phalloidin solution in a moist chamber at room temperature for 30 min |
Applications | When used TRITC Phalloidin staining to examine the presence of F-actin in the amoeba nucleus. No significant amount of F-actin was detected in the nucleus; instead, F-actin formed a cytoplasmic meshwork of filaments and bundles supporting the nucleus |
参考文献: [1]. Lv Y, Chen C, et,al. Regulation of matrix stiffness on the epithelial-mesenchymal transition of breast cancer cells under hypoxia environment. Naturwissenschaften. 2017 Jun;104(5-6):38. doi: 10.1007/s00114-017-1461-9. Epub 2017 Apr 5. PMID: 28382476. [2]. Berdieva M, Bogolyubov D, et,al. Nucleus-associated actin in Amoeba proteus. Eur J Protistol. 2016 Oct;56:191-199. doi: 10.1016/j.ejop.2016.09.002. Epub 2016 Sep 9. PMID: 27684042. | |
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