MGEA5 ELISA Kit (Rat)

Intra-assay Precision (Precision within an assay) : 3 samples with low, middle and high level Meningioma Expressed Antigen 5 (MGEA5) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays) : 3 samples with low, middle and high level Meningioma Expressed Antigen 5 (MGEA5) were tested on 3 different plates, 8 replicates in each plate. CV (%) = SD/meanX100 Intra-Assay: CV<10% Inter-Assay: CV<12%

Beta-hexosaminidase; beta-N-acetylhexosaminidase; bifunctional protein NCOAT; meningioma expressed antigen 5 (hyaluronidase) ; meningioma-expressed antigen 5; Mgea5; N-acetyl-beta-D-glucosaminidase; N-acetyl-beta-glucosaminidase; Ncoat; nuclear cytoplasmic O-GlcNAcase and acetyltransferase; protein O-GlcNAcase; protein O-GlcNAcase-like.

Isoform 1: Cleaves GlcNAc but not GalNAc from O-glycosylated proteins. Can use p-nitrophenyl-beta-GlcNAc and 4-methylumbelliferone-GlcNAc as substrates but not p-nitrophenyl-beta-GalNAc or p-nitrophenyl-alpha-GlcNAc (in vitro) (PubMed:8034696) . Does not bind acetyl-CoA and does not have histone acetyltransferase activity.

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 Meningioma Expressed Antigen 5 (MGEA5) . Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Meningioma Expressed Antigen 5 (MGEA5) . 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 Meningioma Expressed Antigen 5 (MGEA5), 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 Meningioma Expressed Antigen 5 (MGEA5) 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

Component Amount Anti-Mgea5 Microplate 96 Wells (12 x 8 Well Strips) Mgea5 Lyophilized Standard 2 x 100 ng 100X Biotinylated Mgea5 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-Mgea5 Microplate 96 Wells (12 x 8 Well Strips) Anti-Mgea5 Microplate 96 Wells (12 x 8 Well Strips) Mgea5 Lyophilized Standard 2 x 100 ng Mgea5 Lyophilized Standard 2 x 100 ng 100X Biotinylated Mgea5 Detector Antibody 120 uL 100X Biotinylated Mgea5 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 N-Acetyl Beta-D-Glucosaminidase (NAGase) . No significant cross-reactivity or interference between N-Acetyl Beta-D-Glucosaminidase (NAGase) and analogues was observed.