MIA ELISA Kit (Rat)

Intra-assay Precision (Precision within an assay) : 3 samples with low, middle and high level Melanoma Inhibitory Activity Protein 1 (MIA1) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays) : 3 samples with low, middle and high level Melanoma Inhibitory Activity Protein 1 (MIA1) were tested on 3 different plates, 8 replicates in each plate. CV (%) = SD/meanX100 Intra-Assay: CV<10% Inter-Assay: CV<12%

Cartilage-derived retinoic acid-sensitive protein; Cdrap; CD-RAP; melanoma inhibitory activity 1; melanoma inhibitory activity protein; melanoma inhibitory activity/condrocyte-derived retinoic acid sensitive protein homolog (MIA/CD-RAP) ; melanoma-derived growth regulatory protein; melanoma-derived growth regulatory protein-like; Mia1.

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 Melanoma Inhibitory Activity Protein 1 (MIA1) . Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Melanoma Inhibitory Activity Protein 1 (MIA1) . 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 Melanoma Inhibitory Activity Protein 1 (MIA1), 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 Melanoma Inhibitory Activity Protein 1 (MIA1) 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) 84-97 88 EDTA Plasma (n=5) 88-101 95 Heparin Plasma (n=5) 96-103 99 Matrix Recovery range (%) Average (%) Matrix Recovery range (%) Average (%) Serum (n=5) 84-97 88 Serum (n=5) 84-97 88 EDTA Plasma (n=5) 88-101 95 EDTA Plasma (n=5) 88-101 95 Heparin Plasma (n=5) 96-103 99 Heparin Plasma (n=5) 96-103 99

Component Amount Anti-MIA Microplate 96 Wells (12 x 8 Well strips) MIA Lyophilized Standard 2 100X Biotinylated MIA Detector Antibody 1 x 120 uL 100X Avidin-HRP Conjugate 1 x 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 Stop Solution 1 x 6 mL TMB Substrate 1 x 9 mL Component Amount Component Amount Anti-MIA Microplate 96 Wells (12 x 8 Well strips) Anti-MIA Microplate 96 Wells (12 x 8 Well strips) MIA Lyophilized Standard 2 MIA Lyophilized Standard 2 100X Biotinylated MIA Detector Antibody 1 x 120 uL 100X Biotinylated MIA Detector Antibody 1 x 120 uL 100X Avidin-HRP Conjugate 1 x 120 uL 100X Avidin-HRP Conjugate 1 x 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 Stop Solution 1 x 6 mL Stop Solution 1 x 6 mL TMB Substrate 1 x 9 mL TMB Substrate 1 x 9 mL

This assay has high sensitivity and excellent specificity for detection of Melanoma Inhibitory Activity Protein 1 (MIA1) . No significant cross-reactivity or interference between Melanoma Inhibitory Activity Protein 1 (MIA1) and analogues was observed.