Anti-VPS35 antibody (ab10099)
Key features and details
- Goat polyclonal to VPS35
- Suitable for: IHC-P, WB, ICC/IF
- Knockout validated
- 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 -
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.
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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...
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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 |
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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 | (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. |
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. |
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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Immunocytochemistry/Immunofluorescence analysis of paraformaldehyde fixed U2OS cells, permeabilized with 0.15% Triton. Primary incubation 1hr (10µg/mL) followed by Alexa Fluor 488 secondary antibody (2µg/mL), showing cytoplasmic/vesicle staining. The nuclear stain is DAPI (blue).
Negative control: Unimmunized goat IgG (10µg/mL) followed by Alexa Fluor 488 secondary antibody (2µg/mL). -
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. -
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 -
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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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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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. -
Immunohistochemical analysis of paraffin embedded human prostate tissue labeling VPS35 with ab10099 at 8 ug/ml.
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Negative Control showing staining of paraffin embedded Human Prostate, with no primary antibody.
Protocols
Datasheets and documents
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Datasheet download
References (58)
ab10099 has been referenced in 58 publications.
- Giridharan SSP et al. Lipid kinases VPS34 and PIKfyve coordinate a phosphoinositide cascade to regulate retriever-mediated recycling on endosomes. Elife 11:N/A (2022). PubMed: 35040777
- Courtellemont T et al. CROP: a retromer-PROPPIN complex mediating membrane fission in the endo-lysosomal system. EMBO J 41:e109646 (2022). PubMed: 35466426
- Hertel A et al. USP32-regulated LAMTOR1 ubiquitination impacts mTORC1 activation and autophagy induction. Cell Rep 41:111653 (2022). PubMed: 36476874
- 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
- Carosi JM et al. Retromer regulates the lysosomal clearance of MAPT/tau. Autophagy 17:2217-2237 (2021). PubMed: 32960680