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PAM ELISA Kit (Human)

CAT: 0247-OKCD00762-96WSize: 96 WellDry Ice: NoHazardous: No
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CAT#:0247-OKCD00762-96WSize:96 Well
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Reproducibility
Intra-assay Precision (Precision within an assay) : 3 samples with low, middle and high level Peptidylglycine Alpha Amidating Monooxygenase (PAM) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays) : 3 samples with low, middle and high level Peptidylglycine Alpha Amidating Monooxygenase (PAM) were tested on 3 different plates, 8 replicates in each plate. CV (%) = SD/meanX100 Intra-Assay: CV<10% Inter-Assay: CV<12%
CAS Number
7732-18-5
Gene Name
Peptidylglycine alpha-amidating monooxygenase
Gene Aliases
PAL; pancreatic peptidylglycine alpha-amidating monooxygenase; peptidyl alpha-amidating enzyme; peptidyl-alpha-hydroxyglycine alpha-amidating lyase; peptidylamidoglycolate lyase; peptidylglycine 2-hydroxylase; peptidyl-glycine alpha-amidating monooxygenase; peptidylglycine alpha-hydroxylating monooxygenase; PHM.
Gene ID
5066
Accession Number
NP_000910
Reactivity
Homo sapiens|Human
Target
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.
Type
ELISA Kit
Applications
Enzyme-linked Immunosorbent assay-Sandwich
Detection Method
Colorimetric
Assay Principle
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 Peptidylglycine Alpha Amidating Monooxygenase (PAM) . Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Peptidylglycine Alpha Amidating Monooxygenase (PAM) . 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 Peptidylglycine Alpha Amidating Monooxygenase (PAM), 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 Peptidylglycine Alpha Amidating Monooxygenase (PAM) in the samples is then determined by comparing the O.D. of the samples to the standard curve.
Assay Protocol
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
Sample Type
Serum, plasma, cerebrospinal fluid and other biological fluids.
Detection Range
31.2-2,000pg/mL
Recovery
Matrix Recovery Range (%) Average (%) serum (n=5) 97-105 101 EDTA plasma (n=5) 86-96 93 heparin plasma (n=5) 87-95 90 Matrix Recovery Range (%) Average (%) Matrix Recovery Range (%) Average (%) serum (n=5) 97-105 101 serum (n=5) 97-105 101 EDTA plasma (n=5) 86-96 93 EDTA plasma (n=5) 86-96 93 heparin plasma (n=5) 87-95 90 heparin plasma (n=5) 87-95 90
Sensitivity
< 13.5 pg/mL
Reconstitution
2°C to 8°C|-20°C
Components
Component Amount Anti-PAM Microplate 96 Wells (12 x 8 Well Strips) PAM Lyophilized Standard 2 x 2 ng 100X Biotinylated PAM 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-PAM Microplate 96 Wells (12 x 8 Well Strips) Anti-PAM Microplate 96 Wells (12 x 8 Well Strips) PAM Lyophilized Standard 2 x 2 ng PAM Lyophilized Standard 2 x 2 ng 100X Biotinylated PAM Detector Antibody 120 uL 100X Biotinylated PAM 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
Notes
Formerly GWB-KBBLT4
Specificity
This assay has high sensitivity and excellent specificity for detection of Peptidylglycine Alpha Amidating Monooxygenase (PAM) . No significant cross-reactivity or interference between Peptidylglycine Alpha Amidating Monooxygenase (PAM) and analogues was observed.
Protein Length
973
NCBI Gene Symbol
PAM
Protein Name
Peptidyl-glycine alpha-amidating monooxygenase
Gene Name URL
PAM
Nucleotide Accession Number
NM_000919.3

Chemical Information

Water

Water is an oxygen hydride consisting of an oxygen atom that is covalently bonded to two hydrogen atoms. It has a role as an amphiprotic solvent, a mouse metabolite, a Saccharomyces cerevisiae metabolite, a member of greenhouse gas, an Escherichia coli metabolite and a human metabolite. It is an oxygen hydride, a mononuclear parent hydride and an inorganic hydroxy compound. It is a conjugate base of an oxonium. It is a conjugate acid of a hydroxide.

CAS Number7732-18-5
PubChem CID962
IUPAC Nameoxidane
Molecular FormulaH2O
Molecular Weight18.015
XLogP-0.5
Topological Polar Surface Area1
Hydrogen Bond Donor Count1
Hydrogen Bond Acceptor Count1
Rotatable Bond Count0
Heavy Atom Count1
Formal Charge0
Complexity0
SMILES
O
InChI
InChI=1S/H2O/h1H2
InChIKey
XLYOFNOQVPJJNP-UHFFFAOYSA-N
Chemical Structure
2D Structure
2D structure of Water
CAS: 7732-18-5
CID: 962
Formula: H2O
MW: 18.015
Interactive 3D Structure
Loading 3D structure...
View on PubChemRaw 3D data
Computed Properties
PropertyValue
Molecular Weight18.015
XLogP3-0.5
Hydrogen Bond Donor Count1
Hydrogen Bond Acceptor Count1
Rotatable Bond Count0
Exact Mass18.010564683
Monoisotopic Mass18.010564683
Topological Polar Surface Area1 Ų
Heavy Atom Count1
Formal Charge0
Complexity0
Isotope Atom Count0
Defined Atom Stereocenter Count0
Undefined Atom Stereocenter Count0
Defined Bond Stereocenter Count0
Undefined Bond Stereocenter Count0
Covalently-Bonded Unit Count1
Compound Is CanonicalizedYes

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