Anti-VPS35 | Vacuolar protein sorting-associated protein 35B (marker of PVC)

Product no: AS20 4402
AS20 4402 | Clonality: Polyclonal  |  Host: Rabbit |  Reactivity: Arabidopsis thaliana | Marker antibody of pre-vacuolar compartments (PVC)
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  • Product Info
  • Immunogen: Recombinant, His6 tagged VPS35b of Arabidopsis thaliana UniProt: F4I0P8, TAIR: At1g75850
    Host: Rabbit
    Clonality: Polyclonal
    Purity: Total IgG. Protein A purified in PBS, 50% glycerol. Filter sterilized.
    Format: Liquid at 2 mg/ml.
    Quantity: 200 g
    Storage: Store at -20C; once reconstituted make aliquots to avoid repeated freeze-thaw cycles. Please remember to spin the tubes briefly prior to opening them to avoid any losses that might occur from material adhering to the cap or sides of the tube.
    Tested applications: ELISA (ELISA), Immunofluorescence (IF), Immunoprecipitation (IP), Western blot (WB)
    Recommended dilution: assay dependent (ELISA), 1: 400 (IF), 1: 100 (IP), 1: 1000 (WB)
    Expected | apparent MW: 89 | 98 kDa
  • Reactivity
  • Confirmed reactivity: Arabidopsis thaliana
    Predicted reactivity: Species of your interest not listed? Contact us
    Not reactive in: No confirmed exceptions from predicted reactivity are currently known
  • Application Examples
  • Western blot with anti-VPS35 antibodies

    Arabidopsis thaliana 19 day-old seedlings were extracted to a crude extract and separated on 12.5 % SDS-PAGE and blotted to PVDF membrane in wet system. Blot was blocked with 3 % skim milk/TBS-T, 1h/RT with agitation. Blot was incubated in the primary antibody at a dilution of 1: 2000 in TBS-T for 1h/RT. The antibody solution was decanted and the blot was washed 4 times for 10 min in TBS-T at RT with agitation. Blot was incubated in matching secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated) diluted to 1:10 000 in for 1h/RT with agitation. The blot was washed as above and developed with a chemiluminescent detection reagent, following manufacture's recommendations.

    Immunofluorescence using anti-VPS35 antibodies

    Immunofluorescent localisation of VPS35. 
    Tobacco NY-2 cells were transnformed with Arabidopsis thaliana VPS35 (left panel), PEP12 (middle panel, which is a PVC marker)

    Method described in details in: Yamazaki et al. (2008). 

  • Background
  • Background: VPS35 (Vacuolar Protein Sorting-associated protein 35) plays a role in vesicular protein sorting. Component of the membrane-associated retromer complex which is essential in endosome-to-Golgi retrograde transport. Also involved in the efficient sorting of seed storage proteins globulin 12S and albumin 2S. The VPS29-VPS26-VPS35 subcomplex may be involved in recycling of specific cargos from endosome to the plasma membrane. Arabidopsis thaliana has three VPS35 homologs designated VPS35a, VPS35b and VPS35c, and among them,VPS35b plays most important role in these processes. Localised to pre-vacuolar compartments (PVC).
  • Product Citations
  • Selected references: Yamazaki et al. (2008). Arabidopsis VPS35, a retromer component, is required for vacuolar protein sorting and involved in plant growth and leaf senescence. Plant Cell Physiol. 2008 Feb;49(2):142-56. doi: 10.1093/pcp/pcn006. (Immunofluorescence, Immunoprecipitation, Western blot)
  • Protocols
  • Agrisera Western Blot protocol and video tutorials

    Protocols to work with plant and algal protein extracts

    Agrisera Educational Posters Collection
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