Mouse Thrombomodulin ELISA Kit PicoKine®

Thrombomodulin (TM), CD141 or BDCA-3 is an integral membrane protein expressed on the surface of endothelial cells and serves as a cofactor for thrombin. It reduces blood coagulation by converting thrombin to an anticoagulant enzyme from a procoagulant enzyme. Thrombomodulin is also expressed on human mesothelial cell, monocyte and a dendritic cell subset. In humans, thrombomodulin is encoded by the THBD gene. The protein has a molecular mass of 74kDa, and consists of a single chain with 5 distinct domains. Thrombomodulin functions as a cofactor in the thrombin-induced activation of protein C in the anticoagulant pathway by forming a 1:1 stoichiometric complex with thrombin. This raises the speed of protein C activation thousandfold. Thrombomodulin-bound thrombin has procoagulant effect at the same time by inhibiting fibrinolysis by cleaving thrombin-activatable fibrinolysis inhibitor (TAFI, aka carboxypeptidase B2) into its active form. Thrombomodulin is a glycoprotein on the surface of endothelial cells that, in addition to binding thrombin, regulates C3b inactivation by factor I. Mutations in the thrombomodulin gene (THBD) have also been reported to be associated with atypical hemolytic-uremic syndrome (aHUS) .

Lots Lot 1 (pg/ml) Lot 2 (pg/ml) Lot 3 (pg/ml) Lot 4 (pg/ml) Mean (pg/ml) Standard Deviation CV (%) Sample 1 154 144 140 148 146 5.17 3.5% Sample 2 450 488 511 451 475 25.81 5.4% Sample 3 1390 1680 1630 1493 1548 114.16 7.3% 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 1 154 144 140 148 146 5.17 3.5% Sample 1 154 144 140 148 146 5.17 3.5% Sample 2 450 488 511 451 475 25.81 5.4% Sample 2 450 488 511 451 475 25.81 5.4% Sample 3 1390 1680 1630 1493 1548 114.16 7.3% Sample 3 1390 1680 1630 1493 1548 114.16 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) 154 450 1390 169 408 1348 Standard deviation 6.46 33.3 57.4 9.46 38.91 60.66 CV (%) 4.2% 7.4% 4% 5.6% 8.9% 4.5% Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 154 450 1390 169 408 1348 Standard deviation 6.46 33.3 57.4 9.46 38.91 60.66 CV (%) 4.2% 7.4% 4% 5.6% 8.9% 4.5% 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) 154 450 1390 169 408 1348 Mean (pg/ml) 154 450 1390 169 408 1348 Standard deviation 6.46 33.3 57.4 9.46 38.91 60.66 Standard deviation 6.46 33.3 57.4 9.46 38.91 60.66 CV (%) 4.2% 7.4% 4% 5.6% 8.9% 4.5% CV (%) 4.2% 7.4% 4% 5.6% 8.9% 4.5%

Thrombomodulin is a specific endothelial cell receptor that forms a 1:1 stoichiometric complex with thrombin. This complex is responsible for the conversion of protein C to the activated protein C (protein Ca) . Once evolved, protein Ca scissions the activated cofactors of the coagulation mechanism, factor Va and factor VIIIa, and thereby reduces the amount of thrombin generated.

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 62.5 125 250 500 1000 2000 4000 O.D. 0.019 0.068 0.104 0.219 0.38 0.769 1.299 1.957 Concentration (pg/ml) 0 62.5 125 250 500 1000 2000 4000 O.D. 0.019 0.068 0.104 0.219 0.38 0.769 1.299 1.957 Concentration (pg/ml) 0 62.5 125 250 500 1000 2000 4000 Concentration (pg/ml) 0 62.5 125 250 500 1000 2000 4000 O.D. 0.019 0.068 0.104 0.219 0.38 0.769 1.299 1.957 O.D. 0.019 0.068 0.104 0.219 0.38 0.769 1.299 1.957

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.