Overview

  • Product nameAnti-VPS35 antibody
    See all VPS35 primary antibodies
  • Description
    Goat polyclonal to VPS35
  • Tested applicationsIHC-P, WB, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Human
    Predicted to work with: Rat, Cow
  • Immunogen

    Synthetic peptide corresponding to Human VPS35 aa 783-796 (C terminal).
    Sequence:

    C-SPESEGPIYEGLIL


    (Peptide available as ab23181)

  • Positive control
    • WB: Human cerebellum tissue lysate. IHC-P: Human testis tissue. ICC/IF: HEK293 and SHSY5Y cells.
  • General notes


    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.

Properties

  • FormLiquid
  • 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.
  • ClonalityPolyclonal
  • IsotypeIgG
  • Research areas

Associated products

Applications

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.

Application Abreviews Notes
IHC-P Use at an assay dependent concentration.
WB Use a concentration of 0.1 - 0.3 µg/ml. Detects a band of approximately 85 kDa (predicted molecular weight: 91 kDa).Can be blocked with Human VPS35 peptide (ab23181).
ICC/IF Use at an assay dependent concentration.

Target

  • 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
  • Database links
  • Alternative names
    • DKFZp434E1211 antibody
    • DKFZp434P1672 antibody
    • FLJ10752 antibody
    • FLJ13588 antibody
    • FLJ20388 antibody
    • hVPS35 antibody
    • Maternal embryonic 3 antibody
    • Maternal-embryonic 3 antibody
    • MEM 3 antibody
    • MEM3 antibody
    • PARK17 antibody
    • TCCCTA00141 antibody
    • Vacuolar protein sorting 35 (yeast) antibody
    • Vacuolar protein sorting 35 antibody
    • Vacuolar protein sorting 35 homolog antibody
    • Vacuolar protein sorting associated protein 35 antibody
    • Vacuolar protein sorting-associated protein 35 antibody
    • Vesicle protein sorting 35 antibody
    • VPS 35 antibody
    • VPS35 antibody
    • VPS35_HUMAN antibody
    see all

Anti-VPS35 antibody images



  • Predicted band size : 91 kDa

    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 goat anti-rabbit IgG (H + L) and goat anti-mouse IgG (H + L) secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging.

  • ab10099 staining Human Testis by IHC-P. Image A) shows cytoplasmic staining of epithelial cells of the epididymis of the rete testis and image B), some leydig cells. Microwaved antigen retrieval with A) Tris/EDTA buffer pH9 at 3µg/ml and B) Citrate buffer pH6 at 10ug/ml, HRP-staining.
  • 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.

    See Abreview

  • 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.

References for Anti-VPS35 antibody (ab10099)

This product has been referenced in:
  • Da Ros M  et al. Retromer vesicles interact with RNA granules in haploid male germ cells. Mol Cell Endocrinol 401:73-83 (2015). Read more (PubMed: 25486514) »
  • Dhungel N  et al. Parkinson's disease genes VPS35 and EIF4G1 interact genetically and converge on a-synuclein. Neuron 85:76-87 (2015). Read more (PubMed: 25533483) »

See all 13 Publications for this product

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (HeLa)
Specification HeLa
Fixative Paraformaldehyde
Permeabilization Yes - 0.1% Triton X-100
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 22°C
Username

Abcam user community

Verified customer

Submitted Dec 20 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (HeLa)
Loading amount 25 µg
Specification HeLa
Gel Running Conditions Reduced Denaturing (12% Tris-Hepes-SDS gel)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Username

Abcam user community

Verified customer

Submitted Dec 18 2012

Thank you for your phone call. DISCOUNT CODE: xxxx Expiration date: xxx I am very pleased to hear you would like to accept our offer and test ab10099 in mouse. This code will give you 1 free PRIMARY ANTIBODY before the expiration dat...

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