VPS35 | Vacuolar protein sorting-associated protein 35B (marker of PVC)
AS20 4402 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana | Marker antibody of pre-vacuolar compartments (PVC)
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Product Information
Host
Rabbit
Clonality
Polyclonal
Purity
Total IgG. Protein A purified in PBS, 50% glycerol. Filter sterilized.
Format
Liquid at 2 mg/ml.
Quantity
200 ĩg
Storage
Store at -20°C; 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
Application examples
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.
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).
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.
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).
Additional information
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)
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