Lumisterol ELISA Kit

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

Lumisterol is a steroid of interest both because its biosynthesis in FUNGI is a target of ANTIFUNGAL AGENTS, notably AZOLES, and because when it is present in SKIN of animals, ULTRAVIOLET RAYS break a bond to result in ERGOCALCIFEROL.

This assay employs the competitive inhibition enzyme immunoassay technique. A monoclonal antibody specific to Lumisterol (Lum) has been pre-coated onto a microplate. A competitive inhibition reaction is launched between biotin labeled Lumisterol (Lum) and unlabeled Lumisterol (Lum) (Standards or samples) with the pre-coated antibody specific to Lumisterol (Lum) . After incubation the unbound conjugate is washed off. Next, avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. The amount of bound HRP conjugate is reverse proportional to the concentration of Lumisterol (Lum) in the sample. After addition of the substrate solution, the intensity of color developed is reverse proportional to the concentration of Lumisterol (Lum) in the sample.

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) 80-95 86 EDTA plasma (n=5) 92-105 99 heparin plasma (n=5) 80-90 82 Matrix Recovery Range (%) Average (%) Matrix Recovery Range (%) Average (%) serum (n=5) 80-95 86 serum (n=5) 80-95 86 EDTA plasma (n=5) 92-105 99 EDTA plasma (n=5) 92-105 99 heparin plasma (n=5) 80-90 82 heparin plasma (n=5) 80-90 82

Component Amount Anti-Lumisterol Microplate 96 Wells (12 x 8 Well Strips) Lumisterol Lyophilized Standard 2 Lumisterol Biotin Complex 1 100X Avidin/HRP Conjugate 120 uL Standard Diluent 1 x 20 mL Biotin Complex Diluent 1 x 12 mL Avidin/HRP 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 Biotin Complex Reconstitution Buffer 1 x 300 uL Component Amount Component Amount Anti-Lumisterol Microplate 96 Wells (12 x 8 Well Strips) Anti-Lumisterol Microplate 96 Wells (12 x 8 Well Strips) Lumisterol Lyophilized Standard 2 Lumisterol Lyophilized Standard 2 Lumisterol Biotin Complex 1 Lumisterol Biotin Complex 1 100X Avidin/HRP Conjugate 120 uL 100X Avidin/HRP Conjugate 120 uL Standard Diluent 1 x 20 mL Standard Diluent 1 x 20 mL Biotin Complex Diluent 1 x 12 mL Biotin Complex Diluent 1 x 12 mL Avidin/HRP Conjugate Diluent 1 x 12 mL Avidin/HRP 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 Biotin Complex Reconstitution Buffer 1 x 300 uL Biotin Complex Reconstitution Buffer 1 x 300 uL