Customize FPG1 Antibody

Description

Identification and Role of FPG-1 Antibody

The FPG-1 antibody is generated in response to the FPG-1 peptide, a 20-amino-acid sequence derived from dengue virus non-structural protein NS4b. This peptide serves as a conformational epitope recognized by IgG antibodies in sera from convalescent dengue patients (>10 days post-symptom onset) .

Structural context: Antibodies targeting FPG-1 belong to the IgG subclass, characterized by a Y-shaped structure with two antigen-binding Fab regions and an Fc region mediating immune effector functions .
Diagnostic mechanism: The FPG-1 peptide is immobilized on microspheres in a multiplex immunoassay (MPIA) to capture IgG antibodies, enabling high-throughput detection of DENV infection .

Diagnostic Performance of FPG-1 Antibody

FPG-1 antibodies exhibit strong sensitivity in identifying DENV infections, particularly in later stages. Key findings include:

Parameter	FPG-1 Antibody Performance
Sensitivity (C-DENV*)	86%
Sensitivity (A-DENV**)	61.1%
Area Under Curve (AUC)	0.8131
Specificity	>80% (when combined with other peptides)

*C-DENV: Convalescent dengue patients (>10 days post-symptom onset); **A-DENV: Acute dengue patients (≤8 days post-symptom onset).

Multiplex Peptide Immunoassay (MPIA)

The FPG-1 peptide was evaluated alongside 19 other flavivirus-derived peptides using Luminex technology:

Assay design: FPG-1 was conjugated to bovine serum albumin (BSA) and coupled to magnetic beads. IgG binding was measured via fluorescent detection .
Comparative performance: FPG-1 outperformed other peptides in detecting convalescent-phase antibodies, with 86% sensitivity versus 60–65% for alternative markers like PM-22 .

Algorithm-Driven Diagnostics

A Random Forest (RF) model incorporating FPG-1 and other peptides improved diagnostic accuracy:

Combined sensitivity: 88.3% for A-DENV and C-DENV samples .
Specificity optimization: The model achieved >80% specificity while maintaining high sensitivity, reducing false positives in regions with co-circulating flaviviruses .

Technical Validation and Limitations

Cross-reactivity: FPG-1 antibodies show minimal cross-reactivity with Zika or yellow fever virus antibodies, enhancing DENV specificity .
Limitations:
- Reduced sensitivity in acute-phase infections (≤8 days).
- Requires complementary markers (e.g., PM-22) for comprehensive detection .

Broader Implications

The FPG-1 antibody system exemplifies the shift toward peptide-based diagnostics for arboviruses, offering advantages over traditional ELISA methods in scalability and cost. Future research may explore its utility in vaccine efficacy trials or as a prognostic marker for severe dengue .

Product Specs

Buffer

Preservative: 0.03% Proclin 300
Constituents: 50% Glycerol, 0.01M PBS, pH 7.4

Form

Liquid

Lead Time

Made-to-order (14-16 weeks)

Synonyms

FPG1 antibody; FPG2 antibody; At1g52500 antibody; F6D8.28Formamidopyrimidine-DNA glycosylase antibody; Fapy-DNA glycosylase antibody; EC 3.2.2.23 antibody; EC 4.2.99.18 antibody; DNA-(apurinic or apyrimidinic site) lyase FPG1 antibody; Formamidopyrimidine-DNA glycosylase 1 antibody; AtFPG-1 antibody; Formamidopyrimidine-DNA glycosylase 2 antibody; AtFPG-2 antibody; Protein MutM homolog 1 antibody; AtMMH-1 antibody; Protein MutM homolog 2 antibody; AtMMH-2 antibody

Target Names

FPG1

Uniprot No.

O80358

Target Background

Function

FPG1 plays a crucial role in the base excision repair (BER) pathway, a major DNA repair system that removes damaged bases caused by oxidation or mutagenic agents. This enzyme functions as a DNA glycosylase, recognizing and removing damaged bases. Notably, FPG1 exhibits high efficiency in processing 4,6-diamino-5-formamidopyrimidine (FapyA), 2,6-diamino-4-hydroxy-5-formamidopyrimidine (FapyG), and advanced oxidation products of 8-oxoguanine (8-oxoG), including guanidinohydantoin and spiroiminodihydantoin. While it possesses marginal activity towards 8-oxoG, FPG1 demonstrates significant AP (apurinic/apyrimidinic) lyase activity. This activity involves cleaving the DNA backbone through beta-delta elimination, generating a single-strand break at the site of the removed base. This break exhibits both 3'- and 5'-phosphates.

Gene References Into Functions

A notable difference between AthFpg and bacterial Fpg proteins is the absence of the alphaF-beta9/10 loop. This loop has been identified as essential for 8-oxoG recognition. PMID: 22789755

Database Links

KEGG: ath:AT1G52500

STRING: 3702.AT1G52500.2

UniGene: At.11653

Protein Families

FPG family

Subcellular Location

Nucleus.

Tissue Specificity

Expressed in leaves (at protein levels).

Q&A

Here’s a structured collection of FAQs for researchers working with FPG1 antibody in academic contexts, synthesized from peer-reviewed methodologies and antibody validation principles:

How do I validate the specificity of FPG1 antibody in my experimental system?

Methodology:
- Perform knockout/knockdown validation using siRNA or CRISPR-Cas9 in target cell lines. Compare staining intensity between wild-type and knockout cells .
- Use orthogonal validation (e.g., Western blot for protein size confirmation alongside immunofluorescence) .
- Test cross-reactivity with structurally related proteins via peptide blocking assays .

What controls are essential for FPG1 antibody-based assays?

Critical controls:
- Biological controls: Tissue/cells lacking FPG1 expression.
- Technical controls:
  - Isotype controls: Match host species and IgG subclass .
  - Fluorescence minus one (FMO): Omit primary antibody to assess background .
  - Pre-adsorption: Pre-incubate antibody with excess antigen to confirm signal specificity .

How do I optimize FPG1 antibody dilution for flow cytometry?

Titration protocol:
- Test dilutions across a 1:50–1:800 range.
- Calculate signal-to-noise ratio (MFI of positive population / MFI of negative control).
- Select the dilution with the highest ratio without background saturation .

How to resolve contradictory FPG1 localization data between IHC and IF?

Troubleshooting framework:

Factor	IHC	IF
Fixation	Formalin may mask epitopes	Methanol/acetone better for membrane targets
Antigen retrieval	Required for formalin-fixed tissues	Rarely used
Permeabilization	Limited for surface antigens	Critical for intracellular targets

Validate with subcellular fractionation followed by Western blot .

How to design a multiplex panel including FPG1 antibody?

Steps:
- Assign FPG1 to a high-sensitivity channel (e.g., PE, APC).
- Check spillover spread matrix for spectral overlap using compensation beads .
- Use tandem dyes for low-abundance targets to minimize channel competition.

What computational tools assist in predicting FPG1 epitope-antibody interactions?

Tools:
- ABodyBuilder: Models antibody-antigen docking .
- BepiPred-3.0: Predicts linear epitopes using machine learning .
- Validate predictions with alanine scanning mutagenesis .

Antibody Validation Checklist

Parameter	Test	Acceptance Criteria
Specificity	Knockout validation	≥90% signal reduction in KO
Batch consistency	Lot-to-lot comparison	CV <15% in MFI
Cross-reactivity	Peptide blocking assay	≥50% signal inhibition

Data Contradiction Analysis Workflow

Replicate experiments across >3 biological replicates.
Compare results with independent techniques (e.g., RNA-FISH for mRNA vs. antibody for protein).
Perform meta-analysis of published FPG1 interaction databases to identify context-dependent expression patterns .

Key Considerations for Publication

Report antibody clone, catalog number, and validation data (e.g., RRID, KO validation images) .
Disclose batch-specific variability if observed (e.g., differential staining between lots) .

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FPG1 Antibody