PCMT1 ELISA Kit (Human)

Intra-assay Precision (Precision within an assay) : 3 samples with low, middle and high level Protein L-Isoaspartate-O-Methyltransferase (PCMT1) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays) : 3 samples with low, middle and high level Protein L-Isoaspartate-O-Methyltransferase (PCMT1) were tested on 3 different plates, 8 replicates in each plate. CV (%) = SD/meanX100 Intra-Assay: CV<10% Inter-Assay: CV<12%

Epididymis secretory sperm binding protein; L-isoaspartyl protein carboxyl methyltransferase; PIMT; protein L-isoaspartyl/D-aspartyl methyltransferase; protein-beta-aspartate methyltransferase; protein-L-isoaspartate (D-aspartate) O-methyltransferase.

Catalyzes the methyl esterification of L-isoaspartyl and D-aspartyl residues in peptides and proteins that result from spontaneous decomposition of normal L-aspartyl and L-asparaginyl residues. It plays a role in the repair and/or degradation of damaged proteins. Acts on EIF4EBP2, microtubule-associated protein 2, calreticulin, clathrin light chains a and b, Ubiquitin carboxyl-terminal hydrolase isozyme L1, phosphatidylethanolamine-binding protein 1, stathmin, beta-synuclein and alpha-synuclein.

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 Protein L-Isoaspartate-O-Methyltransferase (PCMT1) . Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Protein L-Isoaspartate-O-Methyltransferase (PCMT1) . 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 Protein L-Isoaspartate-O-Methyltransferase (PCMT1), 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 Protein L-Isoaspartate-O-Methyltransferase (PCMT1) 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) 78-95 88 EDTA plasma (n=5) 87-95 91 heparin plasma (n=5) 97-105 101 Matrix Recovery Range (%) Average (%) Matrix Recovery Range (%) Average (%) serum (n=5) 78-95 88 serum (n=5) 78-95 88 EDTA plasma (n=5) 87-95 91 EDTA plasma (n=5) 87-95 91 heparin plasma (n=5) 97-105 101 heparin plasma (n=5) 97-105 101

Component Amount Anti-PCMT1 Microplate 96 Wells (12 x 8 Well Strips) PCMT1 Lyophilized Standard 2 x 2 ng 100X Biotinylated PCMT1 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-PCMT1 Microplate 96 Wells (12 x 8 Well Strips) Anti-PCMT1 Microplate 96 Wells (12 x 8 Well Strips) PCMT1 Lyophilized Standard 2 x 2 ng PCMT1 Lyophilized Standard 2 x 2 ng 100X Biotinylated PCMT1 Detector Antibody 120 uL 100X Biotinylated PCMT1 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 Protein L-Isoaspartate-O-Methyltransferase (PCMT1) . No significant cross-reactivity or interference between Protein L-Isoaspartate-O-Methyltransferase (PCMT1) and analogues was observed.