Overview

  • Product name
  • Description
    Goat polyclonal to VPS35
  • Host species
    Goat
  • Tested applications
    Suitable for: IHC-P, WB, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
    Predicted to work with: 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

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
  • Storage buffer
    pH: 7.30
    Preservative: 0.02% Sodium azide
    Constituents: 99% Tris buffered saline, 0.5% BSA
  • Concentration information loading...
  • Purity
    Immunogen affinity purified
  • Purification notes
    Purified from goat serum by ammonium sulphate precipitation followed by antigen affinity chromatography using the immunizing peptide.
  • Primary antibody 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.
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

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 a concentration of 10 µg/ml.

Target

  • Function
    Essential 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 specificity
    Ubiquitous. Highly expressed in heart, brain, placenta, skeletal muscle, spleen, thymus, testis, ovary, small intestine, kidney and colon.
  • Sequence similarities
    Belongs to the VPS35 family.
  • Cellular localization
    Cytoplasm. 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

Images

  • Immunohistochemical analysis of paraffin embedded Human small intestine staining VPS35 at 5µg/ml.Steamed antigen retrieval with citrate buffer pH 6, AP-staining.
  • Immunohistochemical analysis of paraffin embedded Human Prostate staining VPS35 at 5µg/ml. Steamed antigen retrieval with citrate buffer pH 6, AP-staining.
  • All lanes : Anti-VPS35 antibody (ab10099) at 1 µg/ml

    Lane 1 : Human Brain Cerebellum lysate in RIPA buffer
    Lane 2 : Rat Brain lysate in RIPA buffer

    Lysates/proteins at 35 µg per lane.

    Predicted band size: 91 kDa



    Detected by chemiluminescence.

     

  • 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.
  • 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 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

This product has been referenced in:
  • Vazquez-Sanchez S  et al. VPS35 depletion does not impair presynaptic structure and function. Sci Rep 8:2996 (2018). Read more (PubMed: 29445238) »
  • Miller HE  et al. Actin polymerization in the endosomal pathway, but not on the Coxiella-containing vacuole, is essential for pathogen growth. PLoS Pathog 14:e1007005 (2018). Read more (PubMed: 29668757) »
See all 26 Publications for this product

Customer reviews and Q&As

1-4 of 4 Abreviews or Q&A

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

Abcam user community

Verified customer

Submitted Oct 17 2016

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

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

Abcam user community

Verified customer

Submitted Dec 18 2012

Question
Answer

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