SERPINA12 ELISA Kit (Mouse)

Intra-assay Precision (Precision within an assay) : 3 samples with low, middle and high level Visceral Adipose Tissue Derived Serine Protease Inhibitor (Vaspin) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays) : 3 samples with low, middle and high level Visceral Adipose Tissue Derived Serine Protease Inhibitor (Vaspin) were tested on 3 different plates, 8 replicates in each plate. CV (%) = SD/meanX100 Intra-Assay: CV<10% Inter-Assay: CV<12%

4632419J12Rik; serine (or cysteine) proteinase inhibitor, clade A (alpha-1 antiproteinase, antitrypsin), member 12; serpin A12; vaspin; visceral adipose tissue-derived serine protease inhibitor; visceral adipose-specific SERPIN.

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 Visceral Adipose Tissue Derived Serine Protease Inhibitor (Vaspin) . Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Visceral Adipose Tissue Derived Serine Protease Inhibitor (Vaspin) . 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 Visceral Adipose Tissue Derived Serine Protease Inhibitor (Vaspin), 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 Visceral Adipose Tissue Derived Serine Protease Inhibitor (Vaspin) 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 (%) Serum (n=5) 90-104 98 EDTA Plasma (n=5) 84-104 90 Heparin Plasma (n=5) 94-102 98 Matrix Recovery range (%) Average (%) Matrix Recovery range (%) Average (%) Serum (n=5) 90-104 98 Serum (n=5) 90-104 98 EDTA Plasma (n=5) 84-104 90 EDTA Plasma (n=5) 84-104 90 Heparin Plasma (n=5) 94-102 98 Heparin Plasma (n=5) 94-102 98

Component Amount Anti-SERPINA12 Microplate 96 Wells (12 x 8 Well strips) SERPINA12 Lyophilized Standard 2 100X Biotinylated SERPINA12 Detector Antibody 1 x 120 uL 100X Avidin-HRP Conjugate 1 x 120 uL Standard Diluent 1 x 20 mL Detector Antibody Diluent 1 x 12 mL Conjugate Diluent 1 x 12 mL 30X Wash Buffer 1 x 20 mL Stop Solution 1 x 6 mL TMB Substrate 1 x 9 mL Component Amount Component Amount Anti-SERPINA12 Microplate 96 Wells (12 x 8 Well strips) Anti-SERPINA12 Microplate 96 Wells (12 x 8 Well strips) SERPINA12 Lyophilized Standard 2 SERPINA12 Lyophilized Standard 2 100X Biotinylated SERPINA12 Detector Antibody 1 x 120 uL 100X Biotinylated SERPINA12 Detector Antibody 1 x 120 uL 100X Avidin-HRP Conjugate 1 x 120 uL 100X Avidin-HRP Conjugate 1 x 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 Stop Solution 1 x 6 mL Stop Solution 1 x 6 mL TMB Substrate 1 x 9 mL TMB Substrate 1 x 9 mL

This assay has high sensitivity and excellent specificity for detection of Visceral Adipose Tissue Derived Serine Protease Inhibitor (Vaspin) . No significant cross-reactivity or interference between Visceral Adipose Tissue Derived Serine Protease Inhibitor (Vaspin) and analogues was observed.