Rat Legumain ELISA Kit PicoKine®

Legumain is a protein that in humans is encoded by the LGMN gene. It is mapped to 6q32. This gene encodes a cysteine protease, legumain, that has a strict specificity for hydrolysis of asparaginyl bonds. This enzyme may be involved in the processing of bacterial peptides and endogenous proteins for MHC class II presentation in the lysosomal/endosomal systems. Enzyme activation is triggered by acidic pH and appears to be autocatalytic. Protein expression occurs after monocytes differentiate into dendritic cells. A fully mature, active enzyme is produced following lipopolysaccharide expression in mature dendritic cells. Overexpression of this gene may be associated with the majority of solid tumor types. This gene has a pseudogene on chromosome 13. Several alternatively spliced transcript variants have been described, but the biological validity of only two has been determined. These two variants encode the same isoform.

Lots Lot 1 (pg/ml) Lot 2 (pg/ml) Lot 3 (pg/ml) Lot 4 (pg/ml) Mean (pg/ml) Standard Deviation CV (%) Sample 1 35 39 42 37 38 2.58 6.7% Sample 2 212 225 226 219 220 5.59 2.5% Sample 3 806 792 727 859 796 47.02 5.9% Lots Lot 1 (pg/ml) Lot 2 (pg/ml) Lot 3 (pg/ml) Lot 4 (pg/ml) Mean (pg/ml) Standard Deviation CV (%) Lots Lot 1 (pg/ml) Lot 2 (pg/ml) Lot 3 (pg/ml) Lot 4 (pg/ml) Mean (pg/ml) Standard Deviation CV (%) Sample 1 35 39 42 37 38 2.58 6.7% Sample 1 35 39 42 37 38 2.58 6.7% Sample 2 212 225 226 219 220 5.59 2.5% Sample 2 212 225 226 219 220 5.59 2.5% Sample 3 806 792 727 859 796 47.02 5.9% Sample 3 806 792 727 859 796 47.02 5.9%

Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 35 212 806 52 234 939 Standard deviation 1.72 10.81 41.91 3.43 13.57 68.55 CV (%) 4.9% 5.1% 5.2% 6.6% 5.8% 7.3% Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 35 212 806 52 234 939 Standard deviation 1.72 10.81 41.91 3.43 13.57 68.55 CV (%) 4.9% 5.1% 5.2% 6.6% 5.8% 7.3% Intra-Assay Precision Inter-Assay Precision Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 n 16 16 16 24 24 24 Mean (pg/ml) 35 212 806 52 234 939 Mean (pg/ml) 35 212 806 52 234 939 Standard deviation 1.72 10.81 41.91 3.43 13.57 68.55 Standard deviation 1.72 10.81 41.91 3.43 13.57 68.55 CV (%) 4.9% 5.1% 5.2% 6.6% 5.8% 7.3% CV (%) 4.9% 5.1% 5.2% 6.6% 5.8% 7.3%

Has a strict specificity for hydrolysis of asparaginyl bonds. Can also cleave aspartyl bonds slowly, especially under acidic conditions. Required for normal lysosomal protein degradation in renal proximal tubules. Required for normal degradation of internalized EGFR. Plays a role in the regulation of cell proliferation via its role in EGFR degradation. May be involved in the processing of proteins for MHC class II antigen presentation in the lysosomal/endosomal system.

Description|Quantity Pre-coated 96-well strip microplate|1 Standard|2 vials Biotinylated antibody (100x) |100ul Avidin-Biotin-Peroxidase Complex (100x) |100ul Sample Diluent|30ml Antibody Diluent|12ml Avidin-Biotin-Peroxidase Diluent|12ml Color Developing Reagent (TMB) |10ml Stop Solution|10ml Wash Buffer (25x) |20ml Adhesive plate sealers|4

Store at 4°C for 6 months, at -20°C for 12 months. Avoid multiple freeze-thaw cycles (Ships with gel ice, can store for up to 3 days in room temperature. Freeze upon receiving.)

Concentration (pg/ml) 0 31.2 62.5 125 250 500 1000 2000 O.D. 0.021 0.092 0.15 0.266 0.483 0.838 1.51 2.624 Concentration (pg/ml) 0 31.2 62.5 125 250 500 1000 2000 O.D. 0.021 0.092 0.15 0.266 0.483 0.838 1.51 2.624 Concentration (pg/ml) 0 31.2 62.5 125 250 500 1000 2000 Concentration (pg/ml) 0 31.2 62.5 125 250 500 1000 2000 O.D. 0.021 0.092 0.15 0.266 0.483 0.838 1.51 2.624 O.D. 0.021 0.092 0.15 0.266 0.483 0.838 1.51 2.624

Microplate Reader capable of reading absorbance at 450nm. Incubator. Automated plate washer (optional) . Pipettes and pipette tips capable of precisely dispensing 0.5 μl through 1 ml volumes of aqueous solutions. Multichannel pipettes are recommended for large amount of samples. Deionized or distilled water. 500ml graduated cylinders. Test tubes for dilution.