- Datasheet
- Specific References (23)
- Protocols
Overview
-
Product nameAnti-VPS35 antibody
See all VPS35 primary antibodies -
DescriptionGoat polyclonal to VPS35
-
Host speciesGoat
-
Tested applicationsSuitable for: IHC-P, WB, ICC/IFmore details
-
Species reactivityReacts 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 asab23181) -
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
-
Compatible Secondaries
-
Immunizing Peptide (Blocking)
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
- Entrez Gene: 521864 Cow
- Entrez Gene: 100133770 Human
- Entrez Gene: 55737 Human
- Entrez Gene: 65114 Mouse
- Entrez Gene: 25479 Rat
- Omim: 601501 Human
- SwissProt: Q2HJG5 Cow
- SwissProt: Q96QK1 Human
see all -
Alternative names
- DKFZp434E1211 antibody
- DKFZp434P1672 antibody
- FLJ10752 antibody
see all
Images
-
Western blot - Anti-VPS35 antibody (ab10099)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 kDaDetected by chemiluminescence.
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-VPS35 antibody (ab10099)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. -
Western blot - Anti-VPS35 antibody (ab10099)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. -
Immunocytochemistry/ Immunofluorescence - Anti-VPS35 antibody (ab10099)This image is courtesy of an anonymous AbreviewImmunofluorescence 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. -
Western blot - Anti-VPS35 antibody (ab10099)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 kDaIncubated with the primary antibody for 1 hour.
-
Immunocytochemistry/ Immunofluorescence - Anti-VPS35 antibody (ab10099)Immunocytochemistry/Immunofluorescence analysis of PFA-fixed and saponin permeabilized HEK293 cells labelling VPS35 with ab10099.
-
Immunocytochemistry/ Immunofluorescence - Anti-VPS35 antibody (ab10099)Immunocytochemistry/Immunofluorescence analysis of PFA-fixed and saponin permeabilized SHSY5Y cells labelling VPS35 with ab10099.
Protocols
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) »
Customer reviews and Q&As
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"