SEC13A Antibody
Description
Table 1: Key Properties of SEC13A Antibodies
Immune Regulation
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SEC13 hypomorphic mice showed reduced MHC I/II expression in macrophages and diminished IFN-γ+ CD8+ T cells, while TGF-β levels remained elevated .
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Elevated serum immunoglobulins and altered B-cell subsets (e.g., reduced CD19+CD5-IL-4+ cells) were observed .
Disease Associations
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Alzheimer’s disease: SEC13 interacts with presenilin-1, implicating it in amyloid precursor protein processing .
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Metabolic disorders: SEC13 facilitates CD36 trafficking in hepatocytes, influencing lipid metabolism .
Table 2: Experimental Applications and Protocols
Technical Considerations
Product Specs
Target Background
Q&A
Basic Research Questions
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How to validate SEC13A antibody specificity in immunoprecipitation (IP) and Western blot (WB) assays?
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Methodological approach:
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Use knockout (KO) cell lines or CRISPR-Cas9-edited SEC13A-deficient models as negative controls .
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Validate cross-reactivity with SEC13L1 isoforms via epitope mapping (SEC13A antibody immunogen: residues 250–C-terminus) .
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Combine siRNA-mediated SEC13A knockdown with rescue experiments using overexpression constructs .
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Validation Step Expected Outcome Common Pitfalls KO cell lysate No band in WB Partial KO efficiency IP + mass spec SEC13A-specific interactors Non-specific bead binding -
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What are optimal experimental conditions for studying SEC13A’s dual role in COPII vesicle biogenesis and mTORC1 regulation?
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Design considerations:
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For COPII studies: Use ER stress inducers (e.g., tunicamycin) and monitor COPII vesicle formation via electron microscopy .
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For mTORC1 assays: Deplete amino acids to activate GATOR1 and inhibit mTORC1, then reintroduce leucine to assess SEC13A-dependent reactivation .
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Include controls for off-target effects (e.g., SEC13L1 siRNA) .
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How to resolve contradictions between SEC13A localization data in nuclear pore complexes vs. endoplasmic reticulum?
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Troubleshooting framework:
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Perform subcellular fractionation followed by compartment-specific markers (e.g., Lamin B1 for nucleus, Calnexin for ER) .
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Use immunofluorescence with super-resolution microscopy to distinguish NPC-associated vs. cytoplasmic SEC13A pools .
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Compare results across cell types (e.g., HeLa vs. adipocytes, where SEC13A regulates adipsin secretion) .
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Advanced Research Questions
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How does SEC13A’s GATOR2 complex role influence mTORC1 dynamics in nutrient-starved microenvironments?
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Mechanistic insights:
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Use FRET-based mTORC1 biosensors to quantify real-time activity changes upon SEC13A inhibition .
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Profile ubiquitination status of NPRL2 (GATOR1 subunit) under amino acid deprivation ± SEC13A antibodies .
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Correlate SEC13A expression levels with mTORC1-driven metabolic markers (e.g., p-S6K) in tumor biopsies .
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What computational tools predict SEC13A antibody-antigen binding stability for structural studies?
Data Interpretation Guidelines
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Cross-reactivity alerts: SEC13A antibodies may detect SEC13L1 in tissues with high proteolytic activity (e.g., liver) . Confirm using isoform-specific qPCR.
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Functional redundancy: In SEC13A-depleted cells, monitor compensatory upregulation of COPII components (e.g., SEC23/24) .
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Immune link: SEC13A-associated "Maturation" gene signatures (e.g., CD40, CD86) correlate with adaptive immune activation . Use CyTOF to map SEC13A+ dendritic cell subsets in inflammatory models .
