Inositol Triphosphate ELISA Kit

Intra-assay Precision (Precision within an assay) : 3 samples with low, middle and high level Inositol Triphosphate (IP3) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays) : 3 samples with low, middle and high level Inositol Triphosphate (IP3) were tested on 3 different plates, 8 replicates in each plate. CV (%) = SD/meanX100 Intra-Assay: CV<10% Inter-Assay: CV<12%

Inositol 1,4,5-trisphosphate; 1,4,5-Insp3; D-myo-Inositol 1,4,5-trisphosphate; InsP3; 1D-myo-Inositol 1,4,5-trisphosphate; D-myo-inositol-1,4,5-triphosphate

Intracellular messenger formed by the action of phospholipase C on phosphatidylinositol 4,5-bisphosphate, which is one of the phospholipids that make up the cell membrane. Inositol 1,4,5-trisphosphate is released into the cytoplasm where it releases calcium ions from internal stores within the cell's endoplasmic reticulum. These calcium ions stimulate the activity of B kinase or calmodulin.

This assay employs the competitive inhibition enzyme immunoassay technique. A monoclonal antibody specific to Inositol Triphosphate (IP3) has been pre-coated onto a microplate. A competitive inhibition reaction is launched between biotin labeled Inositol Triphosphate (IP3) and unlabeled Inositol Triphosphate (IP3) (Standards or samples) with the pre-coated antibody specific to Inositol Triphosphate (IP3) . 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 Inositol Triphosphate (IP3) in the sample. After addition of the substrate solution, the intensity of color developed is reverse proportional to the concentration of Inositol Triphosphate (IP3) 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) 78-94 84 EDTA plasma (n=5) 81-91 88 heparin plasma (n=5) 85-103 98 Matrix Recovery Range (%) Average (%) Matrix Recovery Range (%) Average (%) serum (n=5) 78-94 84 serum (n=5) 78-94 84 EDTA plasma (n=5) 81-91 88 EDTA plasma (n=5) 81-91 88 heparin plasma (n=5) 85-103 98 heparin plasma (n=5) 85-103 98

Component Amount Anti-Inositol Triphosphate Microplate 96 Wells (12 x 8 Well Strips) Inositol Triphosphate Lyophilized Standard 2 100X Inositol Triphosphate Biotin Complex 120 uL 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 Component Amount Component Amount Anti-Inositol Triphosphate Microplate 96 Wells (12 x 8 Well Strips) Anti-Inositol Triphosphate Microplate 96 Wells (12 x 8 Well Strips) Inositol Triphosphate Lyophilized Standard 2 Inositol Triphosphate Lyophilized Standard 2 100X Inositol Triphosphate Biotin Complex 120 uL 100X Inositol Triphosphate Biotin Complex 120 uL 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