Application | Comment | Organism |
---|---|---|
diagnostics | far-red luminescence imaging technology to visualize tumor specific antigens on cell surfaces in vitro (human hepato-cellular carcinoma cell line Huh-7) and in living bodies (mice xenografted with human Delta-like protein-positive Huh-7 cells), based on a far-red fluorescent indocyanine derivative conjugated to biotinylated Cypridina luciferase linked to an anti-human Delta-like protein (Dlk-1) monoclonal antibody (embryonic antigens expressed on the surface of many cancer cells) via biotin-avidin interaction | Cypridina noctiluca |
Cloned (Comment) | Organism |
---|---|
expression of human Delta-like protein (Dlk-1) antigen in HEK-293 cells, immunization of mice (BALB/c) with HEK293-Dlk-1 cells or with the expression vector that encodes full-length Dlk-1 cDNA | Cypridina noctiluca |
Protein Variants | Comment | Organism |
---|---|---|
additional information | biotynilation of the enzyme by attachment of an Avi-tag of a 16-residue peptide to the C-terminus of the luciferase, conjugation to the indocyanine derivative HiLyte Fluor 647 hydrazine via the glycol-chains of the enzyme | Cypridina noctiluca |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
200000 | - |
SDS-PAGE, reduced antibody-avidin conjugate of the conjugated enzyme (FBP-IgG) | Cypridina noctiluca |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
Cypridina luciferin + O2 | Cypridina noctiluca | - |
oxidized Cypridina luciferin + CO2 + hv | conjugated enzyme exhibits bimodal bioluminescence spectrum at 460 nm and 675 nm, exhibits higher light intensity in mouse blood than unconjugated enzyme, far-red emission peak dominates in blood while the 460 nm peak dominates in buffers | ? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Cypridina noctiluca | - |
sea-firefly | - |
Purification (Comment) | Organism |
---|---|
biotinylated enzyme is incubated with 1 microM sodium metaperiodate in 0.1 M sodium acetate buffer (pH 5.2), reaction stopped by glycerol addition, loading onto a size exclusion column, oxidized biotinylated enzyme eluted with the same buffer, pooling of bioluminescent fractions, concentration by ultrafree-0.5 centrifugal filter device, addition of 10 mM HiLyte Fluor 647 hydrazine in 0.1 M sodium acetate buffer (pH 5.2), after incubation loading onto a size-exclusion column, elution with 0.1 M potassium phosphate buffer (pH 7.2) containing 0.15 M NaCl. Treatment of human Delta-like protein (Dlk-1) antibody with 2-mercaptoethylamine (cleaving hinge-region disulfide bonds between heavy chains of antibody molecules) and conjugation of half antibodies to maleimide-activated avidin and purification on a size-exclusion column results in a 200 kDa conjugate | Cypridina noctiluca |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
Cypridina luciferin + O2 | - |
Cypridina noctiluca | oxidized Cypridina luciferin + CO2 + hv | conjugated enzyme exhibits bimodal bioluminescence spectrum at 460 nm and 675 nm, exhibits higher light intensity in mouse blood than unconjugated enzyme, far-red emission peak dominates in blood while the 460 nm peak dominates in buffers | ? |
Synonyms | Comment | Organism |
---|---|---|
CLuc | - |
Cypridina noctiluca |
Cypridina luciferase | - |
Cypridina noctiluca |
Fbp | biotinylated Cypridina luciferase conjugated to far-red fluorescent indocyanine derivative | Cypridina noctiluca |
FBP-IgG | conjugate of reduced monoclonal antibody to conjugated enzyme FBP | Cypridina noctiluca |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
6.5 | 8 | no difference in normalized bioluminescence spectra of conjugated enzyme in 100 mM sodium phosphate buffer (pH 6.5), 100 mM Tris-HCl buffer (pH 7.4) or 100 mM Tris-HCl buffer (pH 8) each containing 0.1 M NaCl | Cypridina noctiluca |