Recombinant Human GST-NEDD8 E1 (APPBP1/UBA3) Protein, CF
Discontinued Product
Recombinant Human GST-NEDD8 E1 (APPBP1/UBA3) Protein, CF Summary
Product Specifications
Contains an N-terminal GST (glutathione S-transferase) tag
86 kDa (GST-APPBP1)
48 kDa (UBA3)
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.
E-312
| Formulation | X mg/ml (X μM) in 50 mM HEPES pH 7.5, 200 mM NaCl, 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: NEDD8 Activating Enzyme (APPBP1/UBA3)
Neural Precursor Cell Expressed Developmentally Downregulated Gene 8 (NEDD8) Activating Enzyme (APPBP1/UBA3) is a heterodimeric NEDD8-activating (E1) enzyme with a predicted molecular weight of 112 kDa. It is responsible for the first step in the conjugation of NEDD8 to protein substrates. The NEDD8 Activating Enzyme heterodimer is composed of a regulatory subunit, Amyloid beta Precursor Protein Binding Protein 1 (APPBP1), and a catalytic subunit, Ubiquitin-like Modifier Activating Enzyme 3 (UBA3). Human APPBP1 is a 534 amino acid (aa) protein with a predicted molecular weight of 60 kDa that is expressed ubiquitously in fetal tissues and in the adult brain (1). APPBP1 is required for UBA3 neddylation activity, regulates enzyme specificity, and is expressed as two isoforms, the full length protein and a second isoform with an alternate N-terminal, aa1-17, sequence (2). APPBP1 has been shown to drive cell cycle progression, and its expression is increased in the hippocampus of Alzheimer's disease brains (3,4). Human UBA3 is a 463 aa protein with a predicted molecular weight of 52 kDa. It is ubiquitously expressed and shares high aa sequence identity with the C-terminal domain of human UBE1 (5). UBA3 contains an ATP-binding domain and an active site cysteine residue, Cys237 in humans, which are both common to E1 enzymes. Like APPBP1, two isoforms of UBA3 have been identified in humans, the full length protein and a truncated isoform, which lacks aa 8-21. UBA3 is required for cell cycle progression and has been shown to downregulate steroid receptor activation (4,6). Neddylation and its associated enzymes have been implicated in the progression of Alzheimer's disease, via neddylation of APP, and cancer via post-translational modification of oncogenes (7,8).
This protein has a GST tag.
- Chow, N. et al. (1996) J Biol Chem 271:11339.
- Walden, H. et al. (2003) Mol Cell 12:1427.
- Chen, Y. et al. (2003) J Cell Biol 163:27.
- Chen, Y. et al. (2000) J Biol Chem 275:8929.
- Gong, L. & E.T. Yeh (1999) J Biol Chem 274:12036.
- Fan, M. et al. (2002) Mol Endocrinol 16:315.
- Chen, Y. et al. (2012) J Cell Mol Med [Epub ahead of print]
- Soucy, T.A. et al. (2010) Genes Cancer 1:708.
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