PLAA ELISA Kit (Human)

Intra-assay Precision (Precision within an assay) : 3 samples with low, middle and high level Phospholipase A2 Activating Protein (PLAP) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays) : 3 samples with low, middle and high level Phospholipase A2 Activating Protein (PLAP) were tested on 3 different plates, 8 replicates in each plate. CV (%) = SD/meanX100 Intra-Assay: CV<10% Inter-Assay: CV<12%

Plays a role in protein ubiquitination, sorting and degradation through its association with VCP (PubMed:27753622) . Involved in ubiquitin-mediated membrane proteins trafficking to late endosomes in an ESCRT-dependent manner, and hence plays a role in synaptic vesicle recycling (By similarity) . May play a role in macroautophagy, regulating for instance the clearance of damaged lysosomes (PubMed:27753622) . Plays a role in cerebellar Purkinje cell development (By similarity) . Positively regulates cytosolic and calcium-independent phospholipase A2 activities in a tumor necrosis factor alpha (TNF-alpha) - or lipopolysaccharide (LPS) -dependent manner, and hence prostaglandin E2 biosynthesis (PubMed:18291623, PubMed:28007986) .

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 Phospholipase A2 Activating Protein (PLAP) . Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Phospholipase A2 Activating Protein (PLAP) . 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 Phospholipase A2 Activating Protein (PLAP), 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 Phospholipase A2 Activating Protein (PLAP) 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) 99-105 102 EDTA Plasma (n=5) 84-97 91 Heparin Plasma (n=5) 81-103 92 Matrix Recovery Range (%) Average (%) Matrix Recovery Range (%) Average (%) Serum (n=5) 99-105 102 Serum (n=5) 99-105 102 EDTA Plasma (n=5) 84-97 91 EDTA Plasma (n=5) 84-97 91 Heparin Plasma (n=5) 81-103 92 Heparin Plasma (n=5) 81-103 92

Component Amount Anti-PLAA Microplate 96 Wells (12 x 8 Well Strips) PLAA Lyophilized Standard 2 x 20 ng 100X Biotinylated PLAA Detector Antibody 120 uL Avidin/HRP Conjugate 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 TMB Substrate 1 x 9 mL Stop Solution 1 x 6 mL Component Amount Component Amount Anti-PLAA Microplate 96 Wells (12 x 8 Well Strips) Anti-PLAA Microplate 96 Wells (12 x 8 Well Strips) PLAA Lyophilized Standard 2 x 20 ng PLAA Lyophilized Standard 2 x 20 ng 100X Biotinylated PLAA Detector Antibody 120 uL 100X Biotinylated PLAA 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 Phospholipase A2 Activating Protein (PLAA) . No significant cross-reactivity or interference between Phospholipase A2 Activating Protein (PLAA) and analogues was observed.