MCCC2 ELISA Kit (Human)

Intra-assay Precision (Precision within an assay) : 3 samples with low, middle and high level Methylcrotonoyl Coenzyme A Carboxylase 2 (MCCC2) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays) : 3 samples with low, middle and high level Methylcrotonoyl Coenzyme A Carboxylase 2 (MCCC2) were tested on 3 different plates, 8 replicates in each plate. CV (%) = SD/meanX100 Intra-Assay: CV<10% Inter-Assay: CV<12%

3-methylcrotonyl-CoA carboxylase 2;3-methylcrotonyl-CoA carboxylase non-biotin-containing subunit;3-methylcrotonyl-CoA:carbon dioxide ligase subunit beta; biotin carboxylase; MCCase subunit beta; MCCB; MCCCbeta; methylcrotonoyl-CoA carboxylase 2 (beta) ; methylcrotonoyl-CoA carboxylase beta chain, mitochondrial; methylcrotonoyl-Coenzyme A carboxylase 2 (beta) ; non-biotin containing subunit of 3-methylcrotonyl-CoA carboxylase; testicular secretory protein Li 29.

Carboxyltransferase subunit of the 3-methylcrotonyl-CoA carboxylase, an enzyme that catalyzes the conversion of 3-methylcrotonyl-CoA to 3-methylglutaconyl-CoA, a critical step for leucine and isovaleric acid catabolism.

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 Methylcrotonoyl Coenzyme A Carboxylase 2 (MCCC2) . Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Methylcrotonoyl Coenzyme A Carboxylase 2 (MCCC2) . 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 Methylcrotonoyl Coenzyme A Carboxylase 2 (MCCC2), 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 Methylcrotonoyl Coenzyme A Carboxylase 2 (MCCC2) 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) 80-89 84 EDTA plasma (n=5) 90-104 101 heparin plasma (n=5) 98-105 102 Matrix Recovery Range (%) Average (%) Matrix Recovery Range (%) Average (%) serum (n=5) 80-89 84 serum (n=5) 80-89 84 EDTA plasma (n=5) 90-104 101 EDTA plasma (n=5) 90-104 101 heparin plasma (n=5) 98-105 102 heparin plasma (n=5) 98-105 102

Component Amount Anti-MCCC2 Microplate 96 Wells (12 x 8 Well Strips) MCCC2 Lyophilized Standard 2 x 20 ng 100X Biotinylated MCCC2 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-MCCC2 Microplate 96 Wells (12 x 8 Well Strips) Anti-MCCC2 Microplate 96 Wells (12 x 8 Well Strips) MCCC2 Lyophilized Standard 2 x 20 ng MCCC2 Lyophilized Standard 2 x 20 ng 100X Biotinylated MCCC2 Detector Antibody 120 uL 100X Biotinylated MCCC2 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 Methylcrotonoyl Coenzyme A Carboxylase 2 (MCCC2) . No significant cross-reactivity or interference between Methylcrotonoyl Coenzyme A Carboxylase 2 (MCCC2) and analogues was observed.