Key features and details
- Goat polyclonal to VPS35
- Suitable for: WB, ICC/IF
- Reacts with: Mouse, Human
- Isotype: IgG
Product nameAnti-VPS35 antibody
See all VPS35 primary antibodies
DescriptionGoat polyclonal to VPS35
Tested applicationsSuitable for: WB, ICC/IFmore details
Species reactivityReacts with: Mouse, Human
Predicted to work with: Cow
- WB: Human cerebellum and mouse brain tissue lysate. IHC-P: Human testis tissue. ICC/IF: HEK293, U2OS and SHSY5Y cells.
The maintenance of organelles in eukaryotic cells depends on sorting proteins, which ensure the proper delivery of organelle-specific proteins. In S. cerevisiae, VPS35 has a direct role in the retrieval of vacuolar cargo proteins, suggesting that human VPS35 may have an analogous function in the maintenance of lysosomes.
Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
Storage bufferpH: 7.30
Preservative: 0.02% Sodium azide
Constituents: 99% Tris buffered saline, 0.5% BSA
Concentration information loading...
PurityImmunogen affinity purified
Purification notesPurified from goat serum by ammonium sulphate precipitation followed by antigen affinity chromatography using the immunizing peptide.
Primary antibody notesThe maintenance of organelles in eukaryotic cells depends on sorting proteins, which ensure the proper delivery of organelle-specific proteins. In S. cerevisiae, VPS35 has a direct role in the retrieval of vacuolar cargo proteins, suggesting that human VPS35 may have an analogous function in the maintenance of lysosomes.
Our Abpromise guarantee covers the use of ab10099 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||Use a concentration of 0.1 - 0.3 µg/ml. Detects a band of approximately 85 kDa (predicted molecular weight: 91 kDa).
1 hour primary incubation is recommended for this product.
|ICC/IF||Use a concentration of 10 µg/ml.|
FunctionEssential component of the retromer complex, a complex required to retrieve lysosomal enzyme receptors (IGF2R and M6PR) from endosomes to the trans-Golgi network. Also required to regulate transcytosis of the polymeric immunoglobulin receptor (pIgR-pIgA).
Tissue specificityUbiquitous. Highly expressed in heart, brain, placenta, skeletal muscle, spleen, thymus, testis, ovary, small intestine, kidney and colon.
Sequence similaritiesBelongs to the VPS35 family.
Cellular localizationCytoplasm. Membrane.
- Information by UniProt
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Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: VPS35 knockout HAP1 cell lysate (20 µg)
Lane 3: NIH3T3 cell lysate (20 µg)
Lane 4: A549 cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab10099 observed at 85 kDa. Red - loading control, ab181602, observed at 37 kDa.
ab10099 was shown to specifically react with VPS35 when VPS35 knockout samples were used. Wild-type and VPS35 knockout samples were subjected to SDS-PAGE. ab10099 and ab181602 (loading control to GAPDH) were diluted to 1 µg/ml and 1/1000 respectively and incubated overnight at 4°C. Blots were developed with IRDye® 800CW Donkey anti-Goat IgG (H + L) and Donkey Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216779) secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging.
Immunofluorescence analysis of paraformaldehyde fixed U2OS cells, permeabilized with 0.15% Triton. Primary incubation of ab10099 1hr (10 μg/ml) followed by Alexa Fluor 488 secondary antibody (2 μg/ml), showing cytoplasmic staining. The nuclear stain is DAPI (blue). Negative control: Unimmunized goat IgG (10 μg/ml) followed by Alexa Fluor 488 secondary antibody (2 μg/ml).
All lanes : Anti-VPS35 antibody (ab10099) at 1 µg/ml
Lane 1 : Human Cerebellum
Lane 2 : Mouse brain
Lysates/proteins at 35 µg per lane.
Predicted band size: 91 kDa
Immunofluorescence analysis of HeLa cells, staining VPS35 with ab10099.
Cells were fixed with paraformaldehyde, permeabilized with 0.1% Triton X-100 and blocked with 10% serum for 1 hour at 22°C. Samples were incubated with primary antibody (1/150 in 10% Serum in PBS) for 16 hours at 4°C. An AlexaFluor®488-conjugated donkey anti-goat polyclonal IgG (1/300) was used as the secondary antibody.
Anti-VPS35 antibody (ab10099) at 0.1 µg/ml + Human cerebellum tissue lysate in RIPA buffer at 35 µg
Developed using the ECL technique.
Predicted band size: 91 kDa
Incubated with the primary antibody for 1 hour.
Immunocytochemistry/Immunofluorescence analysis of PFA-fixed and saponin permeabilized HEK293 cells labelling VPS35 with ab10099.
Immunocytochemistry/Immunofluorescence analysis of PFA-fixed and saponin permeabilized SHSY5Y cells labelling VPS35 with ab10099.
ab10099 has been referenced in 39 publications.
- Li JG et al. A pharmacological chaperone improves memory by reducing Aß and tau neuropathology in a mouse model with plaques and tangles. Mol Neurodegener 15:1 (2020). PubMed: 31964406
- Wu KY et al. Ependymal Vps35 Promotes Ependymal Cell Differentiation and Survival, Suppresses Microglial Activation, and Prevents Neonatal Hydrocephalus. J Neurosci 40:3862-3879 (2020). PubMed: 32291328
- Pietilä M et al. SORLA regulates endosomal trafficking and oncogenic fitness of HER2. Nat Commun 10:2340 (2019). PubMed: 31138794
- Li JG et al. Full recovery of the Alzheimer's disease phenotype by gain of function of vacuolar protein sorting 35. Mol Psychiatry N/A:N/A (2019). PubMed: 30733594
- Sapmaz A et al. USP32 regulates late endosomal transport and recycling through deubiquitylation of Rab7. Nat Commun 10:1454 (2019). PubMed: 30926795