A2ML1 ELISA Kit (Human)

Intra-assay Precision (Precision within an assay) : 3 samples with low, middle and high level Alpha-2-Macroglobulin Like Protein 1 (a2ML1) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays) : 3 samples with low, middle and high level Alpha-2-Macroglobulin Like Protein 1 (a2ML1) were tested on 3 different plates, 8 replicates in each plate. CV (%) = SD/meanX100 Intra-Assay: CV<10% Inter-Assay: CV<12%

Alpha-2-macroglobulin-like protein 1; C3 and PZP-like alpha-2-macroglobulin domain-containing protein 9; C3 and PZP-like, alpha-2-macroglobulin domain containing 9; CPAMD9; OMS; p170.

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 (By similarity) . Displays inhibitory activity against chymotrypsin, papain, thermolysin, subtilisin A and, to a lesser extent, elastase but not trypsin. May play an important role during desquamation by inhibiting extracellular proteases.

The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Alpha-2-Macroglobulin Like Protein 1 (a2ML1) . Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Alpha-2-Macroglobulin Like Protein 1 (a2ML1) . Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Alpha-2-Macroglobulin Like Protein 1 (a2ML1), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm +/- 10nm. The concentration of Alpha-2-Macroglobulin Like Protein 1 (a2ML1) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Reconstitution & Storage Instructions Western Blotting/Immunoblotting (WB/IB) Protocol Immunohistochemistry (IHC) Protocol Immunocytochemistry (ICC) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Blocking Peptide Competition Protocol (BPCP) Immunoprecipitation (IP) Protocol Antibody Array (AA) Protocol Reconstitution & Storage Instructions Reconstitution & Storage Instructions Western Blotting/Immunoblotting (WB/IB) Protocol Western Blotting/Immunoblotting (WB/IB) Protocol Immunohistochemistry (IHC) Protocol Immunohistochemistry (IHC) Protocol Immunocytochemistry (ICC) Protocol Immunocytochemistry (ICC) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Blocking Peptide Competition Protocol (BPCP) Blocking Peptide Competition Protocol (BPCP) Immunoprecipitation (IP) Protocol Immunoprecipitation (IP) Protocol Antibody Array (AA) Protocol Antibody Array (AA) Protocol

Matrix Recovery Range (%) Average (%) _x005F serum (n=5) 83-103 98 _x005F EDTA plasma (n=5) 99-105 102 _x005F heparin plasma (n=5) 79-91 85 _x005F Matrix Recovery Range (%) Average (%) Matrix Recovery Range (%) Average (%) serum (n=5) 83-103 98 serum (n=5) 83-103 98 EDTA plasma (n=5) 99-105 102 EDTA plasma (n=5) 99-105 102 heparin plasma (n=5) 79-91 85 heparin plasma (n=5) 79-91 85

Component Amount _x005F Anti-A2ML1 Microplate 96 Wells (12 x 8 Well Strips) _x005F A2ML1 Lyophilized Standard 2 x 50 ng _x005F 100X Biotinylated A2ML1 Detector Antibody 120 uL _x005F Avidin/HRP Conjugate 120 uL _x005F Standard Diluent 1 x 20 mL _x005F Detector Antibody Diluent 1 x 12 mL _x005F Conjugate Diluent 1 x 12 mL _x005F 30X Wash Buffer 1 x 20 mL _x005F TMB Substrate 1 x 9 mL _x005F Stop Solution 1 x 6 mL _x005F Component Amount Component Amount Anti-A2ML1 Microplate 96 Wells (12 x 8 Well Strips) Anti-A2ML1 Microplate 96 Wells (12 x 8 Well Strips) A2ML1 Lyophilized Standard 2 x 50 ng A2ML1 Lyophilized Standard 2 x 50 ng 100X Biotinylated A2ML1 Detector Antibody 120 uL 100X Biotinylated A2ML1 Detector Antibody 120 uL Avidin/HRP Conjugate 120 uL Avidin/HRP Conjugate 120 uL Standard Diluent 1 x 20 mL Standard Diluent 1 x 20 mL Detector Antibody Diluent 1 x 12 mL Detector Antibody Diluent 1 x 12 mL Conjugate Diluent 1 x 12 mL Conjugate Diluent 1 x 12 mL 30X Wash Buffer 1 x 20 mL 30X Wash Buffer 1 x 20 mL TMB Substrate 1 x 9 mL TMB Substrate 1 x 9 mL Stop Solution 1 x 6 mL Stop Solution 1 x 6 mL

This assay has high sensitivity and excellent specificity for detection of Alpha-2-Macroglobulin Like Protein 1 (a2ML1) ._x005F No significant cross-reactivity or interference between Alpha-2-Macroglobulin Like Protein 1 (a2ML1) and analogues was observed.