VPS28 Antibody, FITC conjugated

Introduction to VPS28 Antibody, FITC Conjugated

The VPS28 Antibody, FITC conjugated is a fluorescently labeled immunological reagent designed for detecting the VPS28 protein, a core subunit of the ESCRT-I complex. FITC (fluorescein isothiocyanate) conjugation enables visualization of VPS28 localization via fluorescence microscopy, making it indispensable for studying endosomal sorting, viral replication, and cellular trafficking mechanisms. This antibody is widely used in immunofluorescence (IF), immunocytochemistry (ICC), and Western blot (WB) applications, as detailed in research protocols and product specifications .

Applications in Research

The VPS28 Antibody, FITC conjugated, is optimized for fluorescence-based techniques to study VPS28’s role in cellular processes and pathogen interactions. Key applications include:

Immunofluorescence (IF) and Immunocytochemistry (ICC)

• Localization Studies: Visualize VPS28 within endosomal compartments or viral replication complexes.

• Co-localization Experiments: Track interactions between VPS28 and viral proteins (e.g., FMDV 3A, dsRNA) .

• Experimental Workflow:

  1. Fix and permeabilize cells.

  2. Block non-specific binding.

  3. Incubate with primary antibody (dilution determined experimentally) .

  4. Detect with FITC-conjugated secondary antibody (if required).

Western Blot (WB)

• Protein Expression Analysis: Quantify VPS28 levels in lysates, particularly in viral infection models .

• Degradation Studies: Monitor VPS28 downregulation by pathogens like FMDV .

Functional Validation

• Knockdown/Overexpression Experiments: Assess VPS28’s role in viral replication or cellular pathways .

Research Findings and Functional Insights

The VPS28 Antibody, FITC conjugated, has been instrumental in elucidating VPS28’s antiviral and cellular functions:

Antiviral Activity Against FMDV

1. Inhibition of Viral Replication:

   • VPS28 destabilizes FMDV replication complexes (RCs) by associating with the 3A protein and degrading structural proteins (VP0, VP1, VP3) .

   • Overexpression of VPS28 reduces dsRNA levels and viral titers, while knockdown enhances replication .

2. Viral Countermeasures:

   • FMDV degrades VPS28 via:

     • Ubiquitin-proteasome pathway: Mediated by 2B and 3A proteins recruiting E3 ligase TRIM21 .

     • Autophagy: 3Cpro degrades VPS28 via protease activity .

   • Both N-terminal and C-terminal domains of VPS28 are required for antiviral activity .

Cellular Trafficking and ESCRT-I Function

• Endosomal Sorting: VPS28, as part of the ESCRT-I complex (VPS23, VPS28, VPS37, MVB12), facilitates protein sorting into multivesicular bodies .

• Imaging ESCRT-I Dynamics: FITC-conjugated antibodies enable real-time tracking of ESCRT-I localization during membrane remodeling .

Considerations for Experimental Use