Octa-dimethyl-ubiquitin: (CH3)2-MQIFVK*TLTGK*TITLEVEPSDTIENV K*AK*IQDK*EGIPPDQQRLIFAGK*QLEDGR TLSDYNIQK*ESTLHLVLRLRGG-OH where K* = (ε-dimethyl)lysyl.
Product Details
| MW: | ~8.7kDa |
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| Source: | Native bovine. |
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| UniProt ID: | P0CG53 |
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| Formulation: | Lyophilized in HPLC-grade water. |
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| Purity: | ≥95% (SDS-PAGE) |
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| Appearance: | White solid. |
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| Application Notes: | Simplifies kinetics for E1-catalyzed active thioester formation.
Typical concentrations for non-rate limiting support of in vitro conjugation reactions lie in the range 200μM to 1mM depending upon individual conditions. |
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| Solubility: | Soluble and stable in DMSO and aqueous buffers up to 100 mg/mL. The solid will dissolve with gentle vortexing at the desired concentration. |
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| Shipping: | Blue Ice |
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| Long Term Storage: | -20°C |
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| Use/Stability: | Store solid cold and dry at -20C; stable for at least one year. Store solutions at -20C for up to three months. The protein is stable to multiple freeze-thaw cycles. |
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| Scientific Background: | Reductive methylation of the seven internal lysine ε-amino groups as well as the α-amino group to give, in this case, the octa-dimethyl derivative renders ubiquitin unable to form multi-ubiquitin chains via lysine linkages. However, the ability of this methylated ubiquitin to form an active C-terminal thioester is preserved and it may be transferred by ubiquitin-activating and conjugating enzymes (E1 and E2s) to lysine residues within substrate proteins to yield monoubiquitinylated products. |
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| Regulatory Status: | RUO - Research Use Only |
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Mass spectrum analysis by electrospray ionization mass spectrometry, (see image) indicates the octa-dimethylated ubiquitin molecular weight at 8790 Da (± 1.6 Da), doubly charged species present at 6671.7 Da. This is fully consistent with the anticipated molecular weight, given that for parent ubiquitin a molecular weight of 8565.4 Da (± 1.6 Da) is observed under identical conditions. There is no un-derivatized ubiquitin present. The signal observed at 8675.3 Da is attributable to a minor contaminant in the starting ubiquitin and is not due to the presence of octa-monomethylated ubiquitin. All other minor signals observed are at background level and are not indicative of protein artifacts.
SDS-PAGE Analysis: Lane 1: MW Marker, Lane 2: 0.5 μg, Lane 3: 1.0 μg, Lane 4: 2.0 μg, Lane 5: 5.0 μg of Purified Ubiquitin (methylated) (bovine), (native) (Prod. No. BML-UW8555).
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