GBF3 Antibody
Description
Definition and Overview
GB3 Antibody refers to monoclonal or polyclonal antibodies specific to globotriaosylceramide (Gb3), a neutral glycosphingolipid expressed on cell membranes. Key characteristics include:
Biochemical Characteristics
GB3 Antibody binds to distinct epitopes of Gb3, enabling its use in diagnostic and research contexts:
Key Features
-
Antigen Recognition:
-
Molecular Weight (MW):
Clinical Applications and Research Findings
GB3 Antibody serves as a diagnostic and prognostic tool in specific diseases:
Fabry Disease Cardiomyopathy (FDCM)
-
Mechanism: Anti-Gb3 antibodies mark autoimmune myocardial inflammation in FDCM, potentially guiding therapy .
Cancer Research
-
Burkitt Lymphoma: GB3 (CD77) is a tumor-associated antigen, with antibodies aiding in diagnosis via flow cytometry .
-
Synthesis: Gb3 is synthesized by A4GALT (α4-galactosyltransferase), with polyclonal A4GALT antibodies used to study enzymatic activity .
Production
| Type | Host/Clone | Purification | Applications |
|---|---|---|---|
| Monoclonal | Lewis Rat (IgM, 38.13) | Protein-A affinity chromatography | Flow cytometry, IHC |
| Polyclonal (A4GALT) | Rabbit | Antigen affinity purification | Western blot, ELISA |
Detection Methods
-
ELISA: In-house assays (e.g., FSL-Gb3-coated plates) measure circulating anti-Gb3 levels, with a cut-off of P/N > 2.56 for myocarditis detection .
-
Immunohistochemistry: Identifies Gb3 in tissue biopsies, aiding in FDCM diagnosis .
Table 1: GB3 Antibody Applications
| Application | Method | Purpose |
|---|---|---|
| Flow Cytometry | PE/FITC-conjugated anti-GB3 | Detecting Gb3 on cell surfaces |
| Immunohistochemistry | Frozen section staining | Diagnosing FDCM or Burkitt lymphoma |
| ELISA | Anti-Gb3 IgM detection | Monitoring autoimmune FDCM |
Table 2: GB3 Antibody Performance in FDCM Studies
| Metric | Value | Patient Group |
|---|---|---|
| Sensitivity | 87.5% | FDCM + myocarditis |
| Specificity | 81.1% | FDCM without myocarditis |
| AUC (ROC Curve) | Not reported |
Product Specs
Constituents: 50% Glycerol, 0.01M Phosphate Buffered Saline (PBS), pH 7.4
Target Background
Q&A
Here’s a structured collection of FAQs for researchers working with GBF3 antibodies, based on academic research scenarios and synthesized from peer-reviewed sources:
How to validate GBF3 antibody specificity in plant studies?
-
Method: Perform Western blotting using protein extracts from Arabidopsis thaliana (wild-type vs. GBF3 knockout mutants). Include recombinant GBF3 protein as a positive control.
-
Critical step: Pre-absorb the antibody with excess recombinant GBF3 to confirm signal loss .
-
Data interpretation: A single band at ~70 kDa (predicted molecular weight) confirms specificity .
What experimental designs mitigate cross-reactivity in Brassica species?
How to optimize antibody storage for long-term stability?
-
Protocol:
How to resolve contradictions in antibody performance across assays?
-
Case study: A 2024 analysis of antibody classification errors used confusion matrices and similarity graphs to identify antibody pairs frequently misclassified due to shared epitopes .
-
Strategy:
What computational tools predict GBF3 antibody-antigen interactions?
-
Model: Memory B cell language models (mBLMs) trained on antibody sequences can predict specificity by identifying key residues (e.g., CDR3 motifs) .
How does tissue fixation affect GBF3 antibody binding?
-
Findings:
Fixation Method Nuclear Signal Retention Background Noise Formalin High Moderate Methanol Reduced Low
Quantifying GBF3 expression in heterogeneous tissues
-
Solution: Combine laser capture microdissection with qRT-PCR for cell-type-specific analysis .
-
Data normalization: Use housekeeping genes validated for plant tissues (e.g., ACTIN2 in Arabidopsis) .
Interpreting non-specific bands in Western blots
-
Troubleshooting workflow:
