VPS29 Antibody, HRP conjugated
Description
VPS29 Protein Function and Antibody Applications
VPS29 (Vacuolar Protein Sorting-Associated Protein 29) is a subunit of the retromer complex, mediating cargo recycling from endosomes to the Golgi or plasma membrane. Its roles include:
-
Auxin transport regulation in plants (PIN protein sorting) .
-
Pathogen defense (e.g., Eimeria tenella vaccine development) .
Antibodies targeting VPS29 are employed in:
Table 1: Representative VPS29 Antibodies
Key Observations:
-
Primary Antibodies: Unconjugated VPS29 antibodies are available for diverse species (plants, mammals) and applications.
-
HRP Detection: Conjugation is typically applied to secondary antibodies (e.g., anti-rabbit/goat IgG HRP) for signal amplification in WB and IHC .
Plant Biology: VPS29 in Protein Sorting
In Arabidopsis, VPS29 antibodies (e.g., AS20 4401) were used to demonstrate:
-
PIN2 sorting to vacuoles and PIN1 recycling to the plasma membrane .
-
Seed storage protein sorting (12S globulin, 2S albumin) via Western blot .
Protocol Example:
-
Sample Preparation: Extract Arabidopsis shoot apical meristem.
-
SDS-PAGE: Resolve proteins on 18% gel.
-
Primary Antibody: AS20 4401 (1:2000 dilution).
-
Detection: Anti-rabbit IgG HRP (1:10,000) + chemiluminescence .
Human Cell Biology: Retromer Complex Dynamics
In mammalian cells (e.g., PC-3 prostate cancer line), VPS29 antibodies (ab315334) revealed:
-
Rab7 interaction: VPS29 binds Rab7-GTP via the VPS26-VPS35-VPS29 subcomplex .
-
Functional Knockdown: siRNA-mediated VPS29 depletion disrupts retromer activity, impairing cargo recycling .
Immunoprecipitation Data:
| Sample | VPS29 Level | Rab7 Association |
|---|---|---|
| Control | High | Strong |
| siRNA Knockdown | Low | Weak |
Drosophila Models: Synaptic Transmission
VPS29 mutations in Drosophila caused:
-
Synaptic defects: Elevated Rab7 levels, impaired retromer localization, and disrupted neurotransmitter recycling .
Emerging Applications: Pathogen Research
In Eimeria tenella studies, recombinant VPS29 (rEtVPS29) was used to:
-
Induce immunity: Vaccination with rEtVPS29 reduced oocyst shedding by 32.2% in chickens .
-
Elucidate pathogen sorting: Localization of EtVPS29 in sporozoites via EYFP tagging .
Technical Considerations for HRP-Based Detection
Product Specs
Constituents: 50% Glycerol, 0.01M PBS, pH 7.4
Target Background
- RidL is critical for binding of the L. pneumophila effector to the Vps29 retromer subunit and displacement of the regulator TBC1D5. PMID: 29146912
- The retromer complex is a highly conserved membrane trafficking assembly composed of three proteins - Vps26, Vps29, and Vps35. These proteins are impaired in neurodegenerative diseases. (Review) PMID: 26965691
- This study demonstrated genetic variability of VPS29 in parkinsonism. PMID: 25475142
- Data indicate that vesicular transport proteins VPS35 and VPS29 influence the levels of the other subunit of retromer. PMID: 25937119
- Mutations in the gene composing the retromer cargo recognition subunit are not a common cause of Parkinson's disease. PMID: 24684791
- Conclusion is that VPS29 is a metal ion-independent, rigid scaffolding domain, essential but not sufficient for incorporation of retromer into functional endosomal transport assemblies. PMID: 21629666
- Analysis of the phosphodiesterase/nuclease-like fold and two protein-protein interaction sites in human VPS29. PMID: 15788412
- It was demonstrated that recombinant human Vps29 displays in vitro phosphatase activity towards a serine-phosphorylated peptide, containing the acidic-cluster dileucine motif of the cytoplasmatic tail of the CI-M6PR. PMID: 16737443
- These observations indicate that the mammalian retromer complex assembles by sequential association of SNX1/2 and Vps26-Vps29-Vps35 subcomplexes on endosomal membranes and that SNX1 and SNX2 play interchangeable but essential roles. PMID: 17101778
- Crystal structure of a VPS29-VPS35 subcomplex showing how the metallophosphoesterase-fold subunit VPS29 acts as a scaffold for the carboxy-terminal half of VPS35. PMID: 17891154
- Membrane recruitment of the cargo-selective retromer subcomplex VPS35/29/26 is catalyzed by the small GTPase Rab7 and inhibited by the Rab-GAP TBC1D5. PMID: 19531583
