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MuRF1 (rat), (recombinant) (GST-tag)

 
BML-UW0405-0025 25 µg 313.00 USD
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Product Specification

Alternative Name:TRIM63, Tripartite motif-containing protein 63, E3 Ubiquitin-protein ligase TRIM63
 
MW:~67kDa
 
Source:Produced in E. coli BL21 (λDE3).
 
UniProt ID:Q91Z63
 
Formulation:Liquid. In 50mM TRIS, pH 7.5, containing 150mM NaCl and 1mM DTT.
 
Purity:≥95% (SDS-PAGE)
 
Purity Detail:Purified by affinity chromatography.
 
Biological Activity:Recombinant GST-MuRF1 is competent for both auto-ubiquitinylation and ubiquitinylation of substrate proteins in vitro.
 
Application Notes:Uses:
1. Muscle atrophy studies - role of ubiquitinylation.
2. In vitro generation of ubiquitinylated muscle atrophy pathway components.
3. Investigation of MuRF1’s role in muscle wasting process.
4. Control ubiquitin E3 ligase for auto-ubiquitinylation reactions.
 
Shipping:Shipped on Dry Ice
 
Short Term Storage:-80°C
 
Long Term Storage:-80°C
 
Use/Stability:Stable for at least 12 months after receipt when stored at -80°C.
 
Handling:Avoid freeze/thaw cycles. After opening, prepare aliquots and store at -80°C.
 
Scientific Background:In catabolic conditions where proteolysis is increased, two specific genes for skeletal muscle atrophy, MuRf1 (muscle ring finger-1) and MAFbx (muscle atrophy F-box) (or atrogin- 1), which encode ubiquitin (Ub) ligases, are up-regulated. These ligases bind to and mediate ubiquitination of myofibrillar proteins for subsequent degradation during muscle atrophy and are considered as sensitive markers of muscle protein degradation by the Ub-proteasome system. Recombinant GST-MuRF1 is competent for both auto-ubiquitinylation and ubiquitinylation of substrate proteins, such as troponin I, in vitro.
 
bml-uw0405 coomassie
Figure 1: SDS-PAGE image of GST-MuRF1 (Coomassie blue stained). Lane 1: MW markers (top to bottom) 200, 116, 97, 66, 55, 45, 36 kDa. Lane 2: MuRF1 (rat) (recombinant) (GST) (Prod. No. BML-UW0405) (2µg).
bml-uw0405 wb
Figure 2: Western blot showing auto-ubiquitinylation of GST-MuRF1 in the presence of UbcH5a (Prod. No. BML-UW9050) UbE1 (Prod. No. BML-UW9410) and ubiquitin (Prod. No. BML-UW8795), without [1] or with [2] Mg-ATP. Ubiquitinylated MuRF1 detected by (Ub)n-reactive monoclonal antibody FK2-HRP (1:1'000, Prod. No. BML-PW0150)
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bml-uw0405 coomassie bml-uw0405 wb

General Literature References

The involvement of the ubiquitin proteasome system in human skeletal muscle remodelling and atrophy: A.J. Murton, et al.; Biochim. Biophys. Acta 1782, 730 (2008), Abstract;
Ubiquitin ligases MuRF1 and MAFbx in human skeletal muscle atrophy: L. de Palma, et al.; Joint Bone Spine 75, 53 (2008), Abstract;
Certain pairs of ubiquitin-conjugating enzymes (E2s) and ubiquitin-protein ligases (E3s) synthesize nondegradable forked ubiquitin chains containing all possible isopeptide linkages: H.T. Kim, et al.; J. Biol. Chem. 282, 17375 (2007), Abstract;
Atrogin-1/MAFbx and MuRF1 are downregulated in aging-related loss of skeletal muscle: E. Edstrom, et al.; J. Gerontol. A Biol. Sci. Med. Sci. 61, 663 (2006), Abstract;
Ubiquitin-protein ligases in muscle wasting: P.R. Cao, et al.; Int. J. Biochem. Cell Biol. 37, 2088 (2005), Abstract;
Molecular mechanisms modulating muscle mass: D.J. Glass; Trends Mol. Med. 9, 344 (2003), Abstract;
Identification of ubiquitin ligases required for skeletal muscle atrophy: S.C. Bodine, et al.; Science 294, 1704 (2001), Abstract;

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