Product Details
| Alternative Name: | 26S proteasome non-ATPase regulatory subunit 4, Antisecretory factor 1, Multiubiquitin chain-binding protein |
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| Source: | GST-S5a (human Rpn10) is covalently coupled to agarose at a concentration of 0.5mg protein/ml of settled resin. |
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| UniProt ID: | P55036 |
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| Formulation: | Liquid. The resin is supplied as a slurry in 50mM TRIS, pH 7.5, containing 150mM sodium chloride, 50% glycerol, 1mM DTT and 0.01% sodium azide. |
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| Purity: | ≥95% (SDS-PAGE) |
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| Shipping: | Blue Ice Not Frozen |
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| Long Term Storage: | +4°C |
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| Handling: | Do not freeze. |
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| Scientific Background: | Rpn10/S5a, a subunit of the 19S regulator of the 26S proteasome, binds multi-ubiquitinylated proteins containing chains of at least four ubiquitin moieties. Rpn10/S5a-agarose retains this ability and may be used to identify and purify multi-ubiquitinylated proteins. |
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| Protocol: | Human S5a sequence, with an N-terminal fusion of glutathione S-transferase (GST) (Schistosoma japonicum), was expressed in E. coli. After purification and analysis the expressed protein was coupled to CNBr activated-agarose and unbound proteins removed by washing. The loading of protein is determined spectrophotometrically during preparation and is shown to be approximately 0.5mg protein per ml of settled resin.
For use, it is recommended that polyubiquitinylated proteins are bound in TBS or PBS and, after extensive washing, eluted by heating for 5 minutes at 95ºC in Laemmli loading buffer prior to SDS-PAGE/Western blot analysis. Alternatively polyubiquitinylated proteins can be eluted using buffer containing 7M urea. |
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| Regulatory Status: | RUO - Research Use Only |
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Product Literature References
Activity Dependent Protein Degradation Is Critical for the Formation and Stability of Fear Memory in the Amygdala: T.J. Jarome, et al.; PLoS One
6, e24349 (2011),
Abstract;
Hyperubiquitination of proteins in dilated cardiomyopathy: J. Weekes, et al.; Proteomics
3, 208 (2003),
Abstract;
Purification of poly-ubiquitinated proteins by S5a-affinity chromatography: R. Layfield, et al.; Proteomics
1, 773 (2001),
Abstract;
