Recombinant Human His6-MBP-HOIP/RNF31 Catalytic Domain, CF Summary
Recombinant Human HOIP/RNF31 is a RING domain Ubiquitin ligase (E3) and component of the LUBAC complex that functions downstream of a Ubiquitin-activating (E1) enzyme and a Ubiquitin-conjugating (E2) enzyme to conjugate Ubiquitin to substrate proteins. Reaction conditions will need to be optimized for each specific application. We recommend an initial Recombinant Human HOIP/RNF31 concentration of 0.1-0.5 μM.
Product Datasheets
Carrier Free
CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins.Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration.The carrier free version does not contain BSA.
In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard.In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.
E3-240
| Formulation | X mg/ml (X μM) in 50 mM HEPES pH 7.5, 150 mM NaCl, 10% Glycerol, 1 mM TCEP |
| Shipping | The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below. |
| Stability & Storage: | Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
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Reconstitution Calculator
Background: HOIP/RNF31
HOIP (HOIL-1 Interacting Protein, also known as RNF31) is a RING domain ubiquitin E3 ligase and component of the LUBAC complex. LUBAC, a large (approximately 600 kDa) heteromultimeric assembly consisting of at least HOIP, HOIL-1, and Sharpin, conjugates linear polyubiquitin chains to various substrates and plays a key role in NF-kappa B activation and signaling events mediated through TNF alphaand IFN. Both the catalytic activity and specificity for linear ubiquitin chain formation of LUBAC are contained within the C-terminal RBR (RING-IBR-RING) and LDD (linear ubiquitin chain determining domain) domains of HOIP. HOIL-1 and Sharpin are essential for full-length HOIP activity, but not for HOIP truncation mutants consisting of the RBR and LDD domains. This protein contains N-terminal 6-His and MBP tags, and is useful for generating unanchored linear polyubiquitin chains without the requirement for HOIL-1 or Sharpin. HOIP catalytic domain contains aa’s 699-1072.
- Belgnaoui S.M., et al. (2012) Cell Host Microbe 12: 211-222
- Smit J.J, et al. (2012) EMBO J. 31: 3833-3844
- Stieglitz B., et al. (2012) EMBO Rep. 13: 840-846
- Wauer T. and Komander D. (2013) EMBO J. doi:10.1038/emboj.2013.125
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