VPS32.2 Antibody
Description
Overview of VPSAntibody
VPS32.2 Antibody (Catalog: DF12256) is a rabbit polyclonal antibody designed to detect CHMP4B (Charged Multivesicular Body Protein 4B), also known as VPS32.2 or hVps32-2. This antibody is widely used in molecular biology applications to study ESCRT (Endosomal Sorting Complex Required for Transport)-III complex dynamics, membrane remodeling, and cellular processes like endocytosis and cytokinesis .
Key Features
| Parameter | Details |
|---|---|
| Target Protein | CHMP4B (VPS32.2) |
| Reactivity | Human, Mouse, Rat; Predicted for Pig, Zebrafish, Bovine, Sheep, Rabbit, Dog, Chicken, Xenopus |
| Applications | Western Blot (WB), Immunohistochemistry (IHC), Immunofluorescence (IF/ICC) |
| Molecular Weight | 29–33 kDa (observed), 25 kDa (calculated) |
| Uniprot ID | Q9H444 |
Functional Significance of CHMP4B/VPS32.2
CHMP4B is a core subunit of the ESCRT-III complex, critical for membrane scission during processes such as:
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Endosomal sorting: Formation of intraluminal vesicles (ILVs) in multivesicular bodies (MVBs) .
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Pathogen evasion: Modulation of extracellular vesicle (EV) biogenesis in parasites like Trichomonas vaginalis and Entamoeba histolytica .
Mechanistic Insights
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ESCRT-III Assembly: CHMP4B forms spiral filaments on membranes, driven by its positively charged N-terminus binding negatively charged lipids. Oligomerization is essential for membrane scission .
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Pathogen Interactions: In E. histolytica, CHMP4B homologs (EhVps32) localize to phagosomes and cytoplasmic vesicles, facilitating erythrocyte engulfment and membrane remodeling .
Applications of VPSAntibody
The antibody is validated for:
Western Blot (WB)
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Detects denatured CHMP4B in lysates from human, mouse, and rat cells.
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Example: Used to analyze ESCRT-III protein levels in studies of mitotic defects in Drosophila blood progenitors .
Immunohistochemistry (IHC)
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Localizes CHMP4B in tissue sections, aiding in the study of tumor suppression and Notch signaling regulation .
Immunofluorescence (IF/ICC)
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Visualizes CHMP4B in dynamic membrane structures, such as phagosomes or endosomes, in live or fixed cells .
Cross-Reactivity and Specificity
| Species | Reactivity | Predicted Cross-Reactivity |
|---|---|---|
| Human | Confirmed | Pig, Zebrafish, Bovine |
| Mouse | Confirmed | Sheep, Rabbit, Dog |
| Rat | Confirmed | Chicken, Xenopus |
Notes:
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Non-Reactivity: No cross-reactivity observed with unrelated ESCRT subunits (e.g., VPS28, VPS24) .
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Applications: Optimized for WB, IHC, and IF/ICC; not validated for ELISA or flow cytometry .
ESCRT-III Dynamics
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Membrane Remodeling: CHMP4B’s polymerization drives inward budding of membranes, as shown in C. elegans and human cells .
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Disease Models: Knockdown of ESCRT-III components (e.g., VPS32) in Drosophila increases mitotic potential in blood progenitors, linking ESCRT to tissue homeostasis .
Pathogen Studies
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Parasite Adhesion: In T. vaginalis, VPS32 overexpression enhances host cell adherence via EV-mediated cargo transfer .
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Phagocytosis: In E. histolytica, EhVps32 (CHMP4B homolog) forms helicoidal structures during erythrocyte ingestion .
Technical Considerations
| Parameter | Recommendation |
|---|---|
| Sample Preparation | Lysates for WB, fixed/permeabilized cells for IHC/IF, paraffin sections for IHC |
| Optimal Dilution | Determined empirically (typically 1:500–1:1000 for WB; 1:200–1:500 for IF/ICC) |
| Controls | Use preimmune serum or non-specific IgG to confirm specificity |
Product Specs
Constituents: 50% Glycerol, 0.01M PBS, pH 7.4
