|Expression cDNA Clone and Sequence
Recombinant protein was produced with TrueORF clone, RC209052. Click on the TrueORF clone link to view cDNA and protein sequences.|
||Predicted MW:||56 kDa|
|Purity:||> 80% as determined by SDS-PAGE and Coomassie blue staining|
|Mass Spec Validation:
||This protein has been positively validated by MS/MS by Dr. Robert Moritz at The Institute of Systems Biology.
|Concentration:||>50 ug/mL as determined by microplate BCA method|
|Buffer and Storage:||25 mM Tris.HCl, pH 7.3, 100 mM glycine, 10% glycerol.|
||Recombinant protein was captured through anti-DDK affinity column followed by conventional chromatography steps.
||LAP3 activity verified in a biochemical assay: Leucine aminopeptidase 3 (LAP3, TP309052) activity was measured in a fluorescent biochemical assay. LAP3 catalyzes the removal of unsubstituted N-terminal amino acids from various peptides and is most active on leucine. LAP3 activity was measured in a 100 µl reaction mixture containing 1 mM L-leucine 7-amido-4-methyl coumarin (Leu-AMC), 50 mM Tris, pH 8.0, 4 mM MgCl2, and 1 mM MnCl2. Cleavage of leucine from the AMC moiety results in a strong increase in fluorescence intensity. Fluorescence was measured over time with an excitation wavelength of 380 nm and an emission wavelength of 460 nm. The activity of the enzyme in this system remained constant over six hours.
||Arginine and proline metabolismGlutathione metabolismMetabolic pathways