Anti-VPS35 antibody (ab10099)
Key features and details
- Goat polyclonal to VPS35
- Suitable for: IHC-P, WB, ICC/IF
- Reacts with: Mouse, Human
- Isotype: IgG
Overview
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Product name
Anti-VPS35 antibody
See all VPS35 primary antibodies -
Description
Goat polyclonal to VPS35 -
Host species
Goat -
Tested applications
Suitable for: IHC-P, WB, ICC/IFmore details -
Species reactivity
Reacts with: Mouse, Human
Predicted to work with: Rat, Cow
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Immunogen
Synthetic peptide corresponding to Human VPS35 aa 783-796 (C terminal).
Sequence:C-SPESEGPIYEGLIL
(Peptide available asab23181) -
Positive control
- WB: Human and Mouse Brain lysate. HepG2 and HEK-293 whole cell lysates. IHC-P: Human testis tissue. ICC/IF: U2OS, HeLa, HEK293 cells. IHC-P: Human prostate tissue.
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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.
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
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Properties
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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
Associated products
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Compatible Secondaries
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Isotype control
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Recombinant Protein
Applications
The Abpromise guarantee
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 a concentration of 5 - 8 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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| WB | (2) |
Use a concentration of 0.03 - 3 µg/ml. Detects a band of approximately 85 kDa (predicted molecular weight: 91 kDa).
1 hour primary incubation is recommended for this product. |
| ICC/IF | (1) |
Use a concentration of 10 µg/ml.
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| Notes |
|---|
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IHC-P
Use a concentration of 5 - 8 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
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WB
Use a concentration of 0.03 - 3 µg/ml. Detects a band of approximately 85 kDa (predicted molecular weight: 91 kDa). 1 hour primary incubation is recommended for this product. |
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ICC/IF
Use a concentration of 10 µg/ml. |
Target
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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
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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
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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. -
Western blot - Anti-VPS35 antibody (ab10099)All lanes : Anti-VPS35 antibody (ab10099) at 1 µg/ml
Lane 1 : Human Cerebellum
Lane 2 : Mouse brain
Lysates/proteins at 35 µg per lane.
Predicted band size: 91 kDa -
Western blot - Anti-VPS35 antibody (ab10099)All lanes :
Lane 1 : HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate (35µg protein in RIPA buffer).
Lane 2 : HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate
(35µg protein in RIPA buffer).
Predicted band size: 91 kDaDetected by chemiluminescence.
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Immunocytochemistry/ Immunofluorescence - Anti-VPS35 antibody (ab10099)Immunofluorescence analysis of paraformaldehyde fixed U2OS cells, permeabilized with 0.15% Triton. Primary incubation of ab10099 1hr (10 μg/ml) followed by Alexa Fluor 488 secondary antibody (2 μg/ml), showing cytoplasmic staining. The nuclear stain is DAPI (blue). Negative control: Unimmunized goat IgG (10 μg/ml) followed by Alexa Fluor 488 secondary antibody (2 μg/ml).
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Immunocytochemistry/ Immunofluorescence - Anti-VPS35 antibody (ab10099)Immunofluorescence analysis of paraformaldehyde fixed HEK293 cells, permeabilized with 0.15% Triton. Primary incubation 1hr (10 ug/ml) followed by Alexa Fluor 488 secondary antibody (2 ug/ml), showing cytoplasmic/vesicle staining. The nuclear stain is DAPI (blue).
Negative control: Unimmunized goat IgG (10 ug/ml) followed by Alexa Fluor 488 secondary antibody (2 ug/ml).
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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. -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-VPS35 antibody (ab10099)Immunohistochemical analysis of paraffin embedded human prostate tissue labeling VPS35 with ab10099 at 8 ug/ml.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-VPS35 antibody (ab10099)Negative Control showing staining of paraffin embedded Human Prostate, with no primary antibody.
Protocols
Datasheets and documents
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Datasheet download
References (51)
ab10099 has been referenced in 51 publications.
- Saez JJ et al. Retrograde and Anterograde Transport of Lat-Vesicles during the Immunological Synapse Formation: Defining the Finely-Tuned Mechanism. Cells 10:N/A (2021). PubMed: 33572370
- Tan KW et al. JIP4 is recruited by the phosphoinositide-binding protein Phafin2 to promote recycling tubules on macropinosomes. J Cell Sci 134:N/A (2021). PubMed: 34109410
- Simoes S et al. Tau and other proteins found in Alzheimer's disease spinal fluid are linked to retromer-mediated endosomal traffic in mice and humans. Sci Transl Med 12:N/A (2020). PubMed: 33239387
- Li JG et al. A pharmacological chaperone improves memory by reducing Aß and tau neuropathology in a mouse model with plaques and tangles. Mol Neurodegener 15:1 (2020). PubMed: 31964406
- Wu KY et al. Ependymal Vps35 Promotes Ependymal Cell Differentiation and Survival, Suppresses Microglial Activation, and Prevents Neonatal Hydrocephalus. J Neurosci 40:3862-3879 (2020). PubMed: 32291328
