Human A2M/Alpha 2-Macroglobulin ELISA Kit PicoKine®

Alpha-2-macroglobulin, also known as A2M or CPAMD5 is a large plasma protein found in the blood. This gene is mapped to 12p13.31. Alpha-2-macroglobulin is a protease inhibitor and cytokine transporter. It inhibits many proteases, including trypsin, thrombin and collagenase. A2M is implicated in Alzheimer disease (AD) due to its ability to mediate the clearance and degradation of A-beta, the major component of beta-amyloid deposits. This gene is able to inhibit all four classes of proteinases by a unique 'trapping' mechanism. This protein has a peptide stretch, called the 'bait region' which contains specific cleavage sites for different proteinases. When a proteinase cleaves the bait region, a conformational change is induced in the protein which traps the proteinase. The entrapped enzyme remains active against low molecular weight substrates (activity against high molecular weight substrates is greatly reduced) . Following cleavage in the bait region a thioester bond is hydrolyzed and mediates the covalent binding of the protein to the proteinase.

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 1367 1285 1469 1391 1378 65.61 4.7% Sample 2 3876 3671 3260 3321 3532 253.06 7.1% Sample 3 26784 32034 29630 29138 29396 1864.33 6.3% 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 1367 1285 1469 1391 1378 65.61 4.7% Sample 1 1367 1285 1469 1391 1378 65.61 4.7% Sample 2 3876 3671 3260 3321 3532 253.06 7.1% Sample 2 3876 3671 3260 3321 3532 253.06 7.1% Sample 3 26784 32034 29630 29138 29396 1864.33 6.3% Sample 3 26784 32034 29630 29138 29396 1864.33 6.3%

Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 1367 3876 26784 1316 3942 26889 Standard deviation 97.05 236.43 2062.36 103.96 240.46 2151.12 CV (%) 7.1% 6.1% 7.7% 7.9% 6.1% 8% Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 1367 3876 26784 1316 3942 26889 Standard deviation 97.05 236.43 2062.36 103.96 240.46 2151.12 CV (%) 7.1% 6.1% 7.7% 7.9% 6.1% 8% 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) 1367 3876 26784 1316 3942 26889 Mean (pg/ml) 1367 3876 26784 1316 3942 26889 Standard deviation 97.05 236.43 2062.36 103.96 240.46 2151.12 Standard deviation 97.05 236.43 2062.36 103.96 240.46 2151.12 CV (%) 7.1% 6.1% 7.7% 7.9% 6.1% 8% CV (%) 7.1% 6.1% 7.7% 7.9% 6.1% 8%

Is able to inhibit all four classes of proteinases by a unique 'trapping' mechanism. This protein has a peptide stretch, called the 'bait region' which contains specific cleavage sites for different proteinases. When a proteinase cleaves the bait region, a conformational change is induced in the protein which traps the proteinase. The entrapped enzyme remains active against low molecular weight substrates (activity against high molecular weight substrates is greatly reduced) . Following cleavage in the bait region a thioester bond is hydrolyzed and mediates the covalent binding of the protein to the proteinase.

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 (ng/ml) 0 0.625 1.25 2.5 5 10 20 40 O.D. 0.016 0.137 0.249 0.476 0.829 1.395 2.108 2.443 Concentration (ng/ml) 0 0.625 1.25 2.5 5 10 20 40 O.D. 0.016 0.137 0.249 0.476 0.829 1.395 2.108 2.443 Concentration (ng/ml) 0 0.625 1.25 2.5 5 10 20 40 Concentration (ng/ml) 0 0.625 1.25 2.5 5 10 20 40 O.D. 0.016 0.137 0.249 0.476 0.829 1.395 2.108 2.443 O.D. 0.016 0.137 0.249 0.476 0.829 1.395 2.108 2.443

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.