Recombinant human UBA1 

(Ubiquitin-activating enzyme E1)

Catalog # : EPX-011-REP

Source : Human

Expressed in : SF9 cells

Quantity : 5 µg of recombinant human UBA1 at 0.05µg /µl

Background:

Activates ubiquitin by first adenylating its C-terminal glycine residue with ATP, and thereafter linking this residue to the side chain of a cysteine residue in E1, yielding an ubiquitin-E1 thioester and free AMP.

Protein details:

Recombinant human N-terminal His tagged UBA1 was produced in Baculovirus, purified using FPLC and formulated in a storage buffer containing 20 mM Tris pH 7.65, 150 mM NaCl, 1 mM DTT, 10% glycerol. Protein concentration was determined by spectrometry. >95% purity by SDS-PAGE.

Quality control:

Each lot has been evaluated by 10% Tris Glycine SDS-PAGE.

SDS-PAGE gel of recombinant UBA1 (Lane 2).  Lane 1, protein molecular weight marker.

In vitro glycosylase assay:

A  70 bp DNA containing a G/T mismatch was incubated or not with recombinant MBD4 for 30 minutes at 37°C. The reaction products were treated with NaOH and run on PAGE under denaturing conditions. Note that a 35 bp cut product was generated in the presence of MBD4.

Storage:

-80°C

Guarantee:

For research use only. Products guaranteed stable for 2 years from date of receipt when stored properly.

Sequence:

UBA1 (Mw: 118,789 Da.)

MIHHHHHHSSSPLSKKRRVSGPDPKPGSNCSPAQSVLSEVPSVPTNGMAKNGSEADIDEGLY
SRQLYVLGHEAMKRLQTSSVLVSGLRGLGVEIAKNIILGGVKAVTLHDQGTAQWADLSSQFY
LREEDIGKNRAEVSQPRLAELNSYVPVTAYTGPLVEDFLSGFQVVVLTNTPLEDQLRVGEFC
HNRGIKLVVADTRGLFGQLFCDFGEEMILTDSNGEQPLSAMVSMVTKDNPGVVTCLDEARHG
FESGDFVSFSEVQGMVELNGNQPMEIKVLGPYTFSICDTSNFSDYIRGGIVSQVKVPKKISF
KSLVASLAEPDFVVTDFAKFSRPAQLHIGFQALHQFCAQHGRPPRPRNEEDAAELVALAQAV
NARALPAVQQNNLDEDLIRKLAYVAAGDLAPINAFIGGLAAQEVMKACSGKFMPIMQWLYFD
ALECLPEDKEVLTEDKCLQRQNRYDGQVAVFGSDLQEKLGKQKYFLVGAGAIGCELLKNFAM
IGLGCGEGGEIIVTDMDTIEKSNLNRQFLFRPWDVTKLKSDTAAAAVRQMNPHIRVTSHQNR
VGPDTERIYDDDFFQNLDGVANALDNVDARMYMDRRCVYYRKPLLESGTLGTKGNVQVVIPF
LTESYSSSQDPPEKSIPICTLKNFPNAIEHTLQWARDEFEGLFKQPAENVNQYLTDPKFVER
TLRLAGTQPLEVLEAVQRSLVLQRPQTWADCVTWACHHWHTQYSNNIRQLLHNFPPDQLTSS
GAPFWSGPKRCPHPLTFDVNNPLHLDYVMAAANLFAQTYGLTGSQDRAAVATFLQSVQVPEF
TPKSGVKIHVSDQELQSANASVDDSRLEELKATLPSPDKLPGFKMYPIDFEKDDDSNFHMDF
IVAASNLRAENYDIPSADRHKSKLIAGKIIPAIATTTAAVVGLVCLELYKVVQGHRQLDSYK
NGFLNLALPFFGFSEPLAAPRHQYYNQEWTLWDRFEVQGLQPNGEEMTLKQFLDYFKTEHKL
EITMLSQGVSMLYSFFMPAAKLKERLDQPMTEIVSRVSKRKLGRHVRALVLELCCNDESGED
VEVPYVRYTIR