Human PAM ELISA Kit PicoKine®

Peptidyl-glycine alpha-amidating monooxygenase is an enzyme that catalyzes the biosynthesis of many signaling peptides and, in humans, is encoded by the PAM gene. It is mapped to 5q21.1. This gene encodes a multifunctional protein. The encoded preproprotein is proteolytically processed to generate the mature enzyme. This enzyme includes two domains with distinct catalytic activities, a peptidylglycine alpha-hydroxylating monooxygenase (PHM) domain and a peptidyl-alpha-hydroxyglycine alpha-amidating lyase (PAL) domain. These catalytic domains work sequentially to catalyze the conversion of neuroendocrine peptides to active alpha-amidated products. Alternative splicing results in multiple transcript variants, at least one of which encodes an isoform that is proteolytically processed.

Lots Lot 1 (pg/ml) Lot 2 (pg/ml) Lot 3 (pg/ml) Lot 4 (pg/ml) Mean (pg/ml) Standard Deviation CV (%) Sample 1 164 228 291 184 217 4.51 2.1% Sample 2 572 532 446 618 542 39.73 7.3% Sample 3 4574 4967 4068 4322 4483 147.48 3.3% Lots Lot 1 (pg/ml) Lot 2 (pg/ml) Lot 3 (pg/ml) Lot 4 (pg/ml) Mean (pg/ml) Standard Deviation CV (%) Lots Lot 1 (pg/ml) Lot 2 (pg/ml) Lot 3 (pg/ml) Lot 4 (pg/ml) Mean (pg/ml) Standard Deviation CV (%) Sample 1 164 228 291 184 217 4.51 2.1% Sample 1 164 228 291 184 217 4.51 2.1% Sample 2 572 532 446 618 542 39.73 7.3% Sample 2 572 532 446 618 542 39.73 7.3% Sample 3 4574 4967 4068 4322 4483 147.48 3.3% Sample 3 4574 4967 4068 4322 4483 147.48 3.3%

Peptidyl-glycine alpha-amidating monooxygenase; PAM; Peptidylglycine alpha-hydroxylating monooxygenase; PHM; Peptidyl-alpha-hydroxyglycine alpha-amidating lyase; Peptidylamidoglycolate lyase; PAL; PAM

Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 164 572 4574 250 468 3375 Standard deviation 4.1 31.17 116.18 11.65 34.77 146.48 CV (%) 2.5% 5.5% 2.5% 4.7% 7.4% 4.3% Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 164 572 4574 250 468 3375 Standard deviation 4.1 31.17 116.18 11.65 34.77 146.48 CV (%) 2.5% 5.5% 2.5% 4.7% 7.4% 4.3% Intra-Assay Precision Inter-Assay Precision Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 n 16 16 16 24 24 24 Mean (pg/ml) 164 572 4574 250 468 3375 Mean (pg/ml) 164 572 4574 250 468 3375 Standard deviation 4.1 31.17 116.18 11.65 34.77 146.48 Standard deviation 4.1 31.17 116.18 11.65 34.77 146.48 CV (%) 2.5% 5.5% 2.5% 4.7% 7.4% 4.3% CV (%) 2.5% 5.5% 2.5% 4.7% 7.4% 4.3%

Bifunctional enzyme that catalyzes 2 sequential steps in C-terminal alpha-amidation of peptides. The monooxygenase part produces an unstable peptidyl (2-hydroxyglycine) intermediate that is dismutated to glyoxylate and the corresponding desglycine peptide amide by the lyase part. C-terminal amidation of peptides such as neuropeptides is essential for full biological activity.

Description|Quantity Pre-coated 96-well strip microplate|1 Standard|2 vials Biotinylated antibody (100x) |100ul Avidin-Biotin-Peroxidase Complex (100x) |100ul Sample Diluent|30ml Antibody Diluent|12ml Avidin-Biotin-Peroxidase Diluent|12ml Color Developing Reagent (TMB) |10ml Stop Solution|10ml Wash Buffer (25x) |20ml Adhesive plate sealers|4

Store at 4°C for 6 months, at -20°C for 12 months. Avoid multiple freeze-thaw cycles (Ships with gel ice, can store for up to 3 days in room temperature. Freeze upon receiving.)

Concentration (pg/ml) 0 156 312 625 1250 2500 5000 10000 O.D. 0.045 0.119 0.173 0.283 0.494 0.822 1.396 2.23 Concentration (pg/ml) 0 156 312 625 1250 2500 5000 10000 O.D. 0.045 0.119 0.173 0.283 0.494 0.822 1.396 2.23 Concentration (pg/ml) 0 156 312 625 1250 2500 5000 10000 Concentration (pg/ml) 0 156 312 625 1250 2500 5000 10000 O.D. 0.045 0.119 0.173 0.283 0.494 0.822 1.396 2.23 O.D. 0.045 0.119 0.173 0.283 0.494 0.822 1.396 2.23

Microplate Reader capable of reading absorbance at 450nm. Incubator. Automated plate washer (optional) . Pipettes and pipette tips capable of precisely dispensing 0.5 μl through 1 ml volumes of aqueous solutions. Multichannel pipettes are recommended for large amount of samples. Deionized or distilled water. 500ml graduated cylinders. Test tubes for dilution.