PAM Antibody, FITC conjugated
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Description
Composition and Target Specificity
PAM Antibody, FITC conjugated consists of a polyclonal rabbit IgG raised against recombinant human PAM protein (amino acids 338-497) . The FITC fluorophore is covalently attached via primary amine groups (lysine residues) on the antibody surface . Key characteristics include:
Optimization and Quality Control
Effective FITC conjugation requires precise stoichiometric control to balance signal intensity and antigen-binding capacity:
Optimization Parameters
| FITC:Antibody Ratio (µg/mg) | Outcomes | Source |
|---|---|---|
| 10–40 | Low quenching but weak signal | |
| 40–80 | Optimal brightness/specificity balance | |
| >160 | Solubility issues, signal quenching |
Critical steps:
-
Sodium azide removal: Essential to prevent FITC-azide reactions
-
Chromatographic purification: DEAE Sephadex separates under/over-labeled antibodies
-
Functional validation: Parallel testing using cell staining titrations
Binding Affinity vs Labeling Index
A landmark study demonstrated :
| Labeling Index (FITC/IgG) | Antigen Affinity (Relative %) | Non-Specific Staining |
|---|---|---|
| 2:1 | 98% | Minimal |
| 5:1 | 72% | Moderate |
| 8:1 | 41% | Severe |
Higher FITC incorporation reduces binding affinity due to steric hindrance at the antigen-binding site .
Application-Specific Performance
Stability and Handling
Comparative Product Analysis
| Vendor | Catalog # | Price (50 µg) | Validation Data |
|---|---|---|---|
| Qtonics | QA30616 | $190 | ELISA only |
| Biorbyt | orb416881 | $204 | Human reactivity confirmed |
| Novus | NBP2-34075 | $529* | IHC, WB, IF validated |
*Unconjugated antibody price shown for reference
Research Applications
Recent studies utilizing FITC-conjugated PAM antibodies have:
Product Specs
Buffer
Preservative: 0.03% Proclin 300
Constituents: 50% Glycerol, 0.01M PBS, pH 7.4
Constituents: 50% Glycerol, 0.01M PBS, pH 7.4
Form
Liquid
Lead Time
Generally, we can ship your orders within 1-3 business days after receiving them. Delivery timelines may vary depending on the purchase method or location. Please consult your local distributors for specific delivery information.
Synonyms
AMD_HUMAN antibody; PAL antibody; PAM antibody; Pancreatic peptidylglycine alpha amidating monooxygenase antibody; Peptidyl alpha amidating enzyme antibody; Peptidyl alpha hydroxyglycine alpha amidating lyase antibody; Peptidyl-alpha-hydroxyglycine alpha-amidating lyase antibody; Peptidylamidoglycolate lyase antibody; Peptidylglycine 2 hydroxylase antibody; Peptidylglycine alpha amidating monooxygenase antibody; Peptidylglycine alpha hydroxylating monooxygenase antibody; PHM antibody
Target Names
PAM
Uniprot No.
Target Background
Function
Peptidylglycine alpha-amidating monooxygenase (PAM) is a bifunctional enzyme that catalyzes the post-translational modification of inactive peptidylglycine precursors into their corresponding bioactive alpha-amidated peptides. This terminal modification is essential in the biosynthesis of many neural and endocrine peptides. Alpha-amidation involves two sequential reactions, both catalyzed by separate catalytic domains of the enzyme. The first step, catalyzed by the peptidyl alpha-hydroxylating monooxygenase (PHM) domain, is the copper-, ascorbate-, and O2-dependent stereospecific hydroxylation (with S stereochemistry) at the alpha-carbon (C-alpha) of the C-terminal glycine of the peptidylglycine substrate. The second step, catalyzed by the peptidylglycine amidoglycolate lyase (PAL) domain, is the zinc-dependent cleavage of the N-C-alpha bond, producing the alpha-amidated peptide and glyoxylate. Similarly, PAM catalyzes the two-step conversion of an N-fatty acylglycine to a primary fatty acid amide and glyoxylate.
Gene References Into Functions
- A single nucleotide polymorphism (rs13175330) in the PAM gene has been associated with hypertension and insulin resistance in a Korean population. PMID: 29162152
- PAM's ancient ability to localize to ciliary membranes, which release bioactive ectosomes, may be linked to its capacity to accumulate in intralumenal vesicles and exosomes. PMID: 28377049
- Research indicates that His108 and a substrate molecule are involved in the reductive pathway, while His172 and Tyr79 play crucial roles in the catalytic pathway during the copper-centered electron transfer catalyzed by peptidylglycine monooxygenase. PMID: 26982589
- PAM expression is elevated in the secretory pathway of differentiated neurons. PMID: 26879543
- The sensitivity of Peptidylglycine alpha-Amidating Monooxygenase (PAM) to oxygen levels in neuroendocrine cells has been studied. PMID: 26296884
- Two missense variants in PAM, encoding p.Asp563Gly (frequency of 4.98%) and p.Ser539Trp (frequency of 0.65%), have been linked to a moderately higher risk of type 2 diabetes (OR = 1.23, P = 3.9 x 10(-10) and OR = 1.47, P = 1.7 x 10(-5), respectively). PMID: 24464100
- This study details the production of the catalytic core of human peptidylglycine alpha-hydroxylating monooxygenase (hPHMcc) in Escherichia coli, possessing a N-terminal fusion to thioredoxin (Trx). PMID: 22554821
- Data suggests that catalytic inactivation of PHM due to pH changes is accompanied by a structural shift between two protein states, involving a strong Cu-S interaction that does not involve M314. PMID: 22080626
- The nuclear retention of PAM mRNA is lost when expressing La proteins that lack a conserved nuclear retention element, suggesting a direct association between PAM mRNA and La protein in vivo. PMID: 16107699
Database Links
Protein Families
Peptidyl-alpha-hydroxyglycine alpha-amidating lyase family; Copper type II ascorbate-dependent monooxygenase family
Subcellular Location
Cytoplasmic vesicle, secretory vesicle membrane; Single-pass membrane protein.; [Isoform 1]: Membrane; Single-pass type I membrane protein.; [Isoform 2]: Membrane; Single-pass type I membrane protein.; [Isoform 3]: Secreted. Note=Secreted from secretory granules.; [Isoform 4]: Secreted. Note=Secreted from secretory granules.
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