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Ubiquitin (bovine), (native) (methylated)

 
BML-UW8555-0001 1 mg 135.00 USD
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Octa-dimethyl-ubiquitin: (CH3)2-MQIFVK*TLTGK*TITLEVEPSDTIENV K*AK*IQDK*EGIPPDQQRLIFAGK*QLEDGR TLSDYNIQK*ESTLHLVLRLRGG-OH where K* = (ε-dimethyl)lysyl.

Product Specification

MW:~8.7kDa
 
Source:Native bovine.
 
UniProt ID:P0CG53
 
Formulation:Lyophilized in HPLC-grade water.
 
Purity:≥95% (SDS-PAGE)
 
Appearance:White solid.
 
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.
 
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.
 
Shipping:Shipped on Blue Ice
 
Long Term Storage:-20°C
 
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.
 
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.
 
Ubiquitin (bovine), (native) (methylated) Mass Spectrum Analysis
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.
Ubiquitin (bovine), (native) (methylated) SDS-PAGE
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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Ubiquitin (bovine), (native) (methylated) Mass Spectrum Analysis Ubiquitin (bovine), (native) (methylated) SDS-PAGE

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