HEXA ELISA Kit (Human)

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

Beta-hexosaminidase subunit alpha; beta-N-acetylhexosaminidase subunit alpha; hexosaminidase A (alpha polypeptide) ; hexosaminidase subunit A; N-acetyl-beta-glucosaminidase subunit alpha; TSD.

Responsible for the degradation of GM2 gangliosides, and a variety of other molecules containing terminal N-acetyl hexosamines, in the brain and other tissues. The form B is active against certain oligosaccharides. The form S has no measurable 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 Hexosaminidase A Alpha (HEXa) . Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Hexosaminidase A Alpha (HEXa) . 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 Hexosaminidase A Alpha (HEXa), 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 Hexosaminidase A Alpha (HEXa) 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) 94-103 99 EDTA plasma (n=5) 95-105 102 heparin plasma (n=5) 92-101 96 Matrix Recovery Range (%) Average (%) Matrix Recovery Range (%) Average (%) serum (n=5) 94-103 99 serum (n=5) 94-103 99 EDTA plasma (n=5) 95-105 102 EDTA plasma (n=5) 95-105 102 heparin plasma (n=5) 92-101 96 heparin plasma (n=5) 92-101 96

Component Amount Anti-HEXA Microplate 96 Wells (12 x 8 Well Strips) HEXA Lyophilized Standard 2 x 10 ng 100X Biotinylated HEXA 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-HEXA Microplate 96 Wells (12 x 8 Well Strips) Anti-HEXA Microplate 96 Wells (12 x 8 Well Strips) HEXA Lyophilized Standard 2 x 10 ng HEXA Lyophilized Standard 2 x 10 ng 100X Biotinylated HEXA Detector Antibody 120 uL 100X Biotinylated HEXA 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 Hexosaminidase A Alpha (HEXa) . No significant cross-reactivity or interference between Hexosaminidase A Alpha (HEXa) and analogues was observed.