AGL5 Antibody
Description
Contextual Clarification
The designation "AGL5" may stem from nomenclature confusion:
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AGL (amylo-1,6-glucosidase, 4-alpha-glucanotransferase) is a human gene (UniProt: P35573) encoding the glycogen debranching enzyme, critical for glycogen metabolism .
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In autoimmune lipodystrophy (AGL) studies, patient sera labeled "AGL1" to "AGL5" were used to identify autoantibodies against perilipin 1 (PLIN1), a lipid droplet protein .
Thus, "AGL5" likely refers to either:
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A patient identifier in autoimmune lipodystrophy research (e.g., serum sample AGL5) .
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A misinterpretation of AGL-targeting antibodies used in glycogen storage disease (GSD) research .
Anti-AGL Antibodies
Anti-PLIN1 Autoantibodies in AGL
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Role: Autoantibodies against perilipin 1 (PLIN1) disrupt lipid metabolism by blocking ABHD5 binding to PLIN1, leading to adipose tissue loss in autoimmune AGL .
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Key Findings:
Research Applications and Protocols
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Western Blot: AGL antibodies detect ~160 kDa bands in human liver/kidney extracts .
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Immunofluorescence: Used to localize AGL in tissue sections (e.g., adipose, muscle) .
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Disease Models: AGL knockout studies link enzyme deficiency to GSD3, characterized by hepatomegaly and hypoglycemia .
Critical Analysis of "AGL5 Antibody" Terminology
No peer-reviewed studies or commercial products explicitly reference an "AGL5 Antibody." Potential explanations for the term:
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Patient-Specific Autoantibodies: In autoimmune AGL studies, "AGL5" denotes a patient sample, not an antibody .
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Typographical Error: May refer to isoforms of AGL (e.g., isoform 3 in liver/heart) .
Future Directions
Product Specs
Constituents: 50% Glycerol, 0.01M PBS, pH 7.4
Target Background
- Gab1/SHP2/p38MAPK signaling pathway and Ang-2 are essential regulators of thrombin-induced monocyte adhesion and vascular leakage. PMID: 27241812
- Research has revealed new roles for SHP1 and SHP2: they are indispensable for synchronized ovule maturation and subsequent fertilization/seed maturation, seed mucilage production, and exert an inhibitory effect that counteracts the ABS function in controlling formative divisions and cell shape of the inner integument and seed coat layers. PMID: 27776173
- The stk shp1 shp2 ant quadruple mutant ovule lacks integument development, similar to the ant single mutant. PMID: 20041269
- SHP1/2 and TAGL1, despite their distinct molecular functions, regulate similar activities. They are essential for seed dispersal in Arabidopsis and tomato, respectively. PMID: 19880793
- This research identifies a novel function for SHP1 and SHP2 genes in promoting stigma, style, and medial tissue development. PMID: 19900437
- Bisindolylmaleimide I inhibits the FGF2-mediated association of Shp2 tyrosine phosphatase with Frs2 and Gab1. PMID: 17145761
Q&A
What are the key applications of AGL antibodies in academic research?
AGL antibodies are primarily utilized for studying glycogen metabolism and adipocyte biology. In basic research, they enable detection of AGL protein expression via Western blot (WB) and immunohistochemistry (IHC), particularly in human, mouse, and rat heart tissues . Advanced applications include exploring AGL’s role in glycogen storage disease type III (GSD III) through transgenic models . For instance, researchers use AGL antibodies to validate mini-GDE transgene functionality in GSD III models, ensuring proper glycogen remodeling .
How should AGL antibody specificity be validated in experimental settings?
To confirm specificity:
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Positive controls: Use HEK-293 or Jurkat cells expressing endogenous AGL .
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Negative controls: Include AGL-deficient samples or non-specific IgG antibodies.
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Epitope mapping: Cross-reference with published data showing reactivity to the full-length AGL protein (175 kDa) .
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Cross-reactivity testing: Exclude reactivity with PLIN1 or other lipodystrophy-associated proteins (e.g., ABHD5) .
Table 1: Recommended Validation Protocols
What challenges arise when interpreting AGL antibody data in lipodystrophy studies?
Key issues:
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Epitope overlap: Anti-PLIN1 autoantibodies in AGL patients may cross-react with AGL antibodies if epitopes are conserved .
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Heterogeneous expression: AGL protein levels vary between adipose depots, requiring tissue-specific normalization .
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Methodological bias: WB may detect full-length AGL, while IHC requires antigen retrieval (TE buffer pH 9.0 or citrate pH 6.0) .
Resolution strategies:
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Dual detection: Co-stain with PLIN1 antibodies to distinguish adipocyte-specific vs. AGL-specific signals .
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Quantitative WB: Use densitometry software to normalize AGL signals to β-actin or GAPDH .
How do AGL antibodies differ from PLIN1 antibodies in studying acquired generalized lipodystrophy (AGL)?
Key distinction: AGL antibodies track enzyme activity, while PLIN1 antibodies identify autoimmune drivers in AGL .
What experimental designs optimize AGL antibody performance in IHC?
Optimized protocol:
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Fixation: Paraffin-embedded sections with formalin fixation .
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Antigen retrieval: TE buffer (pH 9.0) or citrate (pH 6.0) for 20–30 minutes .
Troubleshooting:
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Weak signal: Increase incubation time (overnight at 4°C).
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Background: Use secondary antibodies with low cross-reactivity (e.g., anti-rabbit IgG HRP) .
How do AGL antibodies inform glycogen storage disease (GSD) research models?
Case study: In GSD III models, AGL antibodies validate transgene efficacy by detecting mini-GDE (debranching enzyme) expression. Researchers use WB to confirm proper glycogen remodeling in liver or muscle tissues .
Key applications:
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Protein quantification: WB detects AGL expression levels in knockout vs. rescued models .
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Localization studies: IHC identifies AGL distribution in lysosomes or cytoplasm .
What are the limitations of using AGL antibodies in cross-species studies?
Challenges:
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Reactivity variance: Observed 165 kDa band in human/mouse/rat, but potential epitope differences in non-mammalian models .
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Isoform specificity: Polyclonal antibodies may detect multiple AGL isoforms, complicating interpretation .
Mitigation:
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Ortholog alignment: Compare AGL sequences between species to predict epitope conservation.
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Species-specific controls: Use non-human samples (e.g., rat heart) as positive controls .
How do AGL antibodies contribute to understanding adipocyte biology in metabolic disorders?
AGL antibodies enable tracking of glycogen storage in adipocytes, linking energy metabolism to lipodystrophy. For example, in PLIN1-negative AGL patients, AGL antibodies may reveal compensatory glycogen regulation mechanisms .
Advanced application:
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Multi-omics integration: Combine AGL WB/IHC data with lipidomics to correlate protein expression with metabolic profiles .
What data analysis strategies address conflicting results between AGL antibody and PLIN1 autoantibody studies?
Scenario: A patient tests negative for PLIN1 autoantibodies but shows AGL upregulation.
Resolution:
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Epitope mapping: Use peptide arrays to confirm AGL antibody specificity vs. PLIN1 cross-reactivity .
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Functional assays: Measure ABHD5 displacement (PLIN1’s ligand) to distinguish autoimmune vs. metabolic drivers .
How can AGL antibody data inform therapeutic strategies for metabolic diseases?
Potential applications:
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Biomarker discovery: Quantify AGL expression in GSD III patients to monitor treatment response .
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drug target validation: Use AGL antibodies to assess glycogen remodeling in response to enzyme replacement therapies .
Future directions: Developing AGL-targeting antibodies for modulating glycogen storage in obesity/diabetes models.
