Gelsolin protein: Homo sapiens recombinant

Gelsolin protein: Homo sapiens recombinant
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Product Uses Include

  • Positive control for studying the activity of F-actin severing and capping proteins
  • Investigation of the effect of actin binding proteins (ABPs) on actin dynamics

Material
Gelsolin protein has been purified from E.coli expressing recombinant 6xHIS-tagged human gelsolin (plasma isoform). Gelsolin belongs to a class of actin severing and capping proteins called class I F-actin capping proteins. Each of these class I proteins contains a series of conserved 125-150 amino acid repeat motifs. Gelsolin is characterized by the presence of six repeated motifs, three of which are actin binding domains. Gelsolin exerts a powerful regulatory role on actin filament length and its activity can be modulated by Ca2+ levels, pH , polyphosphoinositides and post-translational modification. Plasma gelsolin contains an additional 25aa leader sequence compared to the cytoplasmic isoform. Gelsolin is supplied as a white lyophilized powder. After reconstitution with of nanopure water, the protein will be in the following buffer: 10 mM Tris pH 7.5, 10 mM NaCl, 0.1 mM MgCl2, 1% (w/v) sucrose, and 0.1% (w/v) dextran. The lyophilized protein is stable at 4°C desiccated (<10% humidity) for 6 months.

Purity
Protein purity is determined by scanning densitometry of Coomassie Blue stained protein on a 4-20% gradient polyacrylamide gel. Gelsolin protein is >90% pure (see Figure 1) and runs at approximately 95 kDa (due to leader sequence and 6xHIS-tag).

HPG6gel

Figure 1. Gelsolin protein purity determination. A 20 µg sample of HPG5 was separated by electrophoresis in a 4-20% SDS-PAGE system and stained with Coomassie Blue. Protein quantitation was performed using the Precision Red Protein Assay Reagent (Cat.# ADV02).

Biological Activity
The biological activity of gelsolin is determined in an F-actin severing assay. F-actin is incubated with gelsolin and the reaction products are centrifuged at 100,000 x g for 1 h. The proportion of actin in the supernatant (G-actin) versus the pellet (F-actin) is compared to a control reaction without gelsolin. Stringent quality control ensures that gelsolin (2 µg) can solubilize 70-80% of F-actin (5 µg) in five min (see Figure 2)

HPG6_comp

Figure 2. Gelsolin biological activity determination. Sample 1 - Control F-actin without gelsolin. Sample 2 - F-actin with 2µg of gelsolin. Sample 3 - 2µg of gelsolin only. S = supernatant, P = pellet.

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

N. Ostler et al. 2014. Gamma interferon-induced guanylate binding protein 1 is a novel actin cytoskeleton remodeling factor. Mol. Cell. Biol. 34, 196-209.

Y.L. Hu et al. 2013. The value of decreased plasma gelsolin levels in patients with systemic lupus erythematosus and rheumatoid arthritis in diagnosis and disease activity evaluation. Lupus. 22, 1455-1461.

C. Campillo et al. 2013. Unexpected membrane dynamics unveiled by membrane nanotube extrusion. Biophys. J. 104, 1248-1256.

T.-F. Wang et al. 2012. Identification and characterization of the actin-binding motif of phostensin. Int. J. Mol. Sci. 13, 15967-15982.

N. Haverland et al. 2010. Immunoreactivity of anti-gelsolin antibodies: implications for biomarker validation. J. Transl. Med. 8, 137.

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