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当前位置: 首页 > 产品中心 > peptide > Cytoskeleton/Fibronectin (Green fluorescent, HiLyte 488)/5x20 ug/FNR02
商品详细Cytoskeleton/Fibronectin (Green fluorescent, HiLyte 488)/5x20 ug/FNR02
Cytoskeleton/Fibronectin (Green fluorescent, HiLyte 488)/5x20 ug/FNR02
Cytoskeleton/Fibronectin (Green fluorescent, HiLyte 488)/5x20 ug/FNR02
商品编号: FNR02
品牌: Cytoskeleton
市场价: ¥7000.00
美元价: 4200.00
产地: 美国(厂家直采)
公司:
产品分类: 多肽合成
公司分类: peptide
联系Q Q: 3392242852
电话号码: 4000-520-616
电子邮箱: info@ebiomall.com
商品介绍
Details

Product Uses Include

  • Observation of fibronectin matrix assembly and cell adhesion
  • Cell invasion assays (1)
  • FACS analysis

 

MaterialFibronectin is purified from bovine plasma.  Protein purity is determined by scanning densitometry of Coomassie Blue stained protein on a 4-20% polyacrylamide gel.  HiLyte Fluor™ 488 labeled fibronectin is >80% pure (Figure 1). 

The protein is modified to contain covalently linked HiLyte Fluor™ 488 at random surface lysines (2). An activated ester of the fluorochrome is used to label the protein. Labeling stoichiometry is determined by spectroscopic measurement of protein and dye concentrations. Final labeling stoichiometry is 1-3 dyes per protein molecule (Figure 2). HiLyte Fluor™ 488 labeled fibronectin can be detected using a filter set of 350-450nm excitation and 500-550 nm emission.

Fibronectin runs as individual subunits on SDS-PAGE with an apparent molecular weight of 230 kDa.  FNR02 is supplied as an orange lyophilized powder.  Each vial of FNR02 contains 20 µg protein.

PurityPurity is determined by scanning densitometry of proteins on SDS-PAGE gels. Samples are >80% pure. 

FNR02fig1

Figure 1:  HiLyte Fluor™ 488 labeled Fibronectin Purity Determination

Legend: 20 µg of unlabeled fibronectin (Lane 1) and 20 µg of HiLyte Fluor™ 488 labeled fibronectin (Lane 2) was separated by electrophoresis in a 4-20% SDS-PAGE system.  The unlabeled protein was stained with Coomassie Blue and visualized in white light. The HiLyte Fluor™ 488 labeled protein was visualized under UV light, no free dye was observed in the dye front. Protein quantitation was determined with the Precision Red™ Protein Assay Reagent (Cat. # ADV02).  Mark12 molecular weight markers are from Invitrogen

FNR02fig2

Figure 2:   Absorption scan of HiLyte Fluor™ 488 labeled fibronectin in solution

Legend:  FNR02 was diluted with Milli-Q water and its absorbance spectrum was scanned between 250 and 650 nm.  HiLyte Fluor™ 488 labeling stoichiometry was calculated to be 1-3 dyes per fibronectin protein using the absorbancy maximum for  at 527 nm and the Beer-Lambert law.  Dye extinction coefficient when protein bound is 70,000M-1cm-1

References

  1. Artym VV. Et al. 2009. ECM degradation assay for analyzing local cell invasion.  Methods in molecular biology, Extracellular matrix protocols, vol. 522: 211-219.Humana Press. 
  2. Use of this product employs the following patent rights licensed to Cytoskeleton, Inc. from Anaspec, Inc.: (a) the claims of U.S. Patents No. 7,754,893, 7,820,783 and 7,790,394; (b) any claims issuing from U.S. Patent Applications Serial No. 12/804,065, 12/807,268 and 12/925,505; and (c) all patents to be issued pursuant thereto, and all continuations, continuations –in-part, reissues, substitutes, and extensions thereof. The use of this product is limited to the field of use comprising internal use by an end user of this product solely in in vivo and in vitro cell staining or biochemical assay applications, such as IHC, HCS, FACS, in vitro assays of an end user only for scientific R&D purposes. The filed of use of this product explicitly excludes the following actions: (a) generating data from clinical applications in humans and animals; and (b) generating QC or QA data for the validation of health, food or cosmetic products.
About

For product Datasheets and MSDSs please click on the PDF links below. For additional information, click on the FAQs tab above or contact our Technical Support department at tservice@cytoskeleton.com

Citations

Rong, Z. et al. Activation of FAK/Rac1/Cdc42‐GTPase signaling ameliorates impaired microglial migration response to Aβ 42 in triggering receptor expressed on myeloid cells 2 loss‐of‐function murine models. FASEB J. 34, 10984–10997 (2020).

Huang, Y., Yi, X., Kang, C. & Wu, C. Arp2/3-Branched Actin Maintains an Active Pool of GTP-RhoA and Controls RhoA Abundance. Cells 8, (2019).

Werley, C. A. et al. Geometry-dependent functional changes in iPSC-derived cardiomyocytes probed by functional imaging and RNA sequencing. PLoS One 12, e0172671 (2017).

Kim, J., Staunton, J. R. & Tanner, K. Independent Control of Topography for 3D Patterning of the ECM Microenvironment. Adv. Mater. 28, 132–137 (2016).

Kim, J. & Tanner, K. Three‐Dimensional Patterning of the ECM Microenvironment Using Magnetic Nanoparticle Self Assembly. Curr. Protoc. Cell Biol. 70, 25.3.1-25.3.14 (2016).

Stanisavljevic, J. et al. Snail1-expressing fibroblasts in the tumor microenvironment display mechanical properties that support metastasis. Cancer Res. 75, 284–295 (2015).

Torr E.E. et al. 2015. Myofibroblasts exhibit enhanced fibronectin assembly that is intrinsic to their contractile phenotype. J. Biol. Chem. 290, 6951-6961.

Jacob A. et al. 2013. Rab40b regulates MMP2 and MMP9 trafficking during invadopodia formation and breast cancer cell invasion. J. Cell Sci. doi: 10.1242/​jcs.126573.

Lively and Schlichter, 2013. The microglial activation state regulates migration and roles of matrix-dissolving enzymes for invasion. J. Neuroinflammation. 10:75.

Faqs

Question 1: What is the optimal excitation and emission filter settings to visualize the HiLyte Fluor™ 488 fluorescence?

Answer 1: HiLyte Fluor™ 488 labeled-fibronectin can be detected using a filter set of 502 nm excitation and 527 nm emission.

Question 2: What is the labeling stoichiometry?

Answer 2: HiLyte Fluor™ 488 labeling stoichiometry was calculated to be 1-3 dyes per fibronectin protein using the absorbancy maximum for HiLyte 488 at 527 nm and the Beer-Lambert law. Dye extinction coefficient when protein bound is 70,000 M-1cm-1.

If you have any questions concerning this product, please contact our Technical Service department at tservice@cytoskeleton.com

品牌介绍
Cytoskeleton,Inc.成立于1993年。自成立以来,我们一直在不断扩大产品范围。Cytoskeleton,Inc.很高兴为药物筛选,信号转导和细胞骨架研究提供广泛的试剂盒和产品。我们专注于纯化蛋白的生产和易于使用的试剂盒,以研究生化和细胞过程。我们的试剂盒既可用于基础研究或小型筛查的少量样品,也可用于大型筛查的高通量规模除了我们现有的产品外,我们还提供产品系列中微管,微管蛋白,运动蛋白,小G蛋白效应物,GAP,GEF和其他几种蛋白的药物筛选服务。有关更多信息,请参见我们的药物筛选服务页面。如果您想从市场上购买到特定产品,请随时与我们联系。我们在这里为您提供帮助。由于我们的科学家在各自的专业领域都有多年的工作经验,因此我们能够提供高质量的产品。自1993年成立以来,我们以合理的价格生产优质的产品而闻名。