Product nameAnti-VPS35 antibody [EPR11501(B)]
See all VPS35 primary antibodies
DescriptionRabbit monoclonal [EPR11501(B)] to VPS35
Tested applicationsSuitable for: WB, IHC-P, Flow Cyt, ICC/IFmore details
Unsuitable for: IP
Species reactivityReacts with: Mouse, Rat, Human
Synthetic peptide within Human VPS35. The exact sequence is proprietary.
- WB: HeLa, HepG2, Raji and MOLT4 cell lysates. IHC-P: Human kidney and human ovarian carcinoma tissue. ICC/IF: MOLT4 and wild-type HAP1 cells.
Storage instructionsShipped at 4°C. Store at -20°C.
Storage bufferPreservative: 0.01% Sodium azide
Constituents: 9% PBS, 40% Glycerol, 0.05% BSA, 50% Tissue culture supernatant
PurityTissue culture supernatant
Our Abpromise guarantee covers the use of ab157220 in the following tested applications.
|WB||1/10000 - 1/50000. Predicted molecular weight: 92 kDa.|
|IHC-P||1/50 - 1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
|Flow Cyt||1/10 - 1/100.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
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: NIH/3T3 cell lysate (20 µg)
Lane 4: A549 cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab157220 observed at 91 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab157220 was shown to specifically react with VPS35 in wild-type HAP1 cells. No band was observed when VPS35 knockout samples were examined. Wild-type and VPS35 knockout samples were subjected to SDS-PAGE. ab157220 and ab8245 (loading control to GAPDH) were diluted 1/1000 and 1/2000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10,000 dilution for 1 hour at room temperature before imaging.
ab157220 staining VPS35 in wild-type HAP1 cells (top panel) and VPS35 knockout HAP1 cells (bottom panel). The cells were fixed with 4% formaldehyde (10min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab157220 at 1/250 dilution and ab195889 at 1/250 dilution (shown in pseudo colour red) overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to Rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.
All lanes : Anti-VPS35 antibody [EPR11501(B)] (ab157220) at 1/10000 dilution
Lane 1 : HeLa cell lysate
Lane 2 : HepG2 cell lysate
Lane 3 : Raji cell lysate
Lane 4 : MOLT4 cell lysate
Lysates/proteins at 10 µg per lane.
All lanes : Goat anti-rabbit HRP at 1/2000 dilution
Predicted band size: 92 kDa
Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling VPS35 with ab157220 at 1/50 dilution.
Immunohistochemical analysis of paraffin-embedded Human ovarian carcinoma tissue labeling VPS35 with ab157220 at 1/50 dilution.
Flow cytometric analysis of permeabilized MOLT4 cells labeling VPS35 with ab157220 at 1/10 dilution (red) or a rabbit IgG (negative) (green).
This product has been referenced in:
- Mir R et al. The Parkinson's disease VPS35[D620N] mutation enhances LRRK2-mediated Rab protein phosphorylation in mouse and human. Biochem J 475:1861-1883 (2018). Read more (PubMed: 29743203) »
- Ma Q et al. SorCS2-mediated NR2A trafficking regulates motor deficits in Huntington's disease. JCI Insight 2:N/A (2017). WB ; Mouse . Read more (PubMed: 28469074) »