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    vps35-antibody-ab10099.pdf

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Signal Transduction Protein Trafficking Vesicle Transport Regulation
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Anti-VPS35 antibody (ab10099)

  • Datasheet
Reviews (3)Q&A (1)References (51)

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Western blot - Anti-VPS35 antibody (ab10099)
  • Western blot - Anti-VPS35 antibody (ab10099)
  • Western blot - Anti-VPS35 antibody (ab10099)
  • Immunocytochemistry/ Immunofluorescence - Anti-VPS35 antibody (ab10099)
  • Immunocytochemistry/ Immunofluorescence - Anti-VPS35 antibody (ab10099)
  • Immunocytochemistry/ Immunofluorescence - Anti-VPS35 antibody (ab10099)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-VPS35 antibody (ab10099)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - 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

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Recombinant Human VPS35 protein (ab132953)

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Overview

  • 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 as ab23181)
    Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI
  • 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.
  • 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.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

Properties

  • 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

    • Signal Transduction
    • Protein Trafficking
    • Vesicle Transport
    • Regulation
    • Signal Transduction
    • Protein Trafficking
    • Golgi Proteins

Associated products

  • Compatible Secondaries

    • Donkey Anti-Goat IgG H&L (Alexa Fluor® 488) (ab150129)
    • Donkey Anti-Goat IgG H&L (HRP) (ab205723)
  • Isotype control

    • Goat IgG, polyclonal - Isotype Control (ab37373)
  • Recombinant Protein

    • Recombinant Human VPS35 protein (ab132953)

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.
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.
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.
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

  • 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.
  • Target information above from: UniProt accession Q96QK1 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    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
    • SwissProt: Q9EQH3 Mouse
    • Unigene: 454528 Human
    • Unigene: 296520 Mouse
    • Unigene: 391632 Mouse
    see all
  • Alternative names

    • DKFZp434E1211 antibody
    • DKFZp434P1672 antibody
    • FLJ10752 antibody
    • FLJ13588 antibody
    • FLJ20388 antibody
    • hVPS35 antibody
    • Maternal embryonic 3 antibody
    • Maternal-embryonic 3 antibody
    • MEM 3 antibody
    • MEM3 antibody
    • PARK17 antibody
    • TCCCTA00141 antibody
    • Vacuolar protein sorting 35 (yeast) antibody
    • Vacuolar protein sorting 35 antibody
    • Vacuolar protein sorting 35 homolog antibody
    • Vacuolar protein sorting associated protein 35 antibody
    • Vacuolar protein sorting-associated protein 35 antibody
    • Vesicle protein sorting 35 antibody
    • VPS 35 antibody
    • VPS35 antibody
    • VPS35_HUMAN antibody
    see all

Images

  • Western blot - Anti-VPS35 antibody (ab10099)
    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)
    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)
    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 kDa



    Detected by chemiluminescence.

  • Immunocytochemistry/ Immunofluorescence - Anti-VPS35 antibody (ab10099)
    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).

  • Immunocytochemistry/ Immunofluorescence - Anti-VPS35 antibody (ab10099)
    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).

  • Immunocytochemistry/ Immunofluorescence - Anti-VPS35 antibody (ab10099)
    Immunocytochemistry/ Immunofluorescence - Anti-VPS35 antibody (ab10099)This image is courtesy of an anonymous Abreview
    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.

    See Abreview

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-VPS35 antibody (ab10099)
    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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-VPS35 antibody (ab10099)
    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

  • Western blot protocols
  • Immunohistochemistry protocols
  • Immunocytochemistry & immunofluorescence protocols

Click here to view the general protocols

Datasheets and documents

  • Datasheet download

    Download

References (51)

Publishing research using ab10099? Please let us know so that we can cite the reference in this datasheet.

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
View all Publications for this product

Customer reviews and Q&As

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1-4 of 4 Abreviews or Q&A

Western blot abreview for Anti-VPS35 antibody

Good
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (HeLa cells)
Gel Running Conditions
Reduced Denaturing
Loading amount
25 µg
Specification
HeLa cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Oct 17 2016

Immunocytochemistry/ Immunofluorescence abreview for Anti-VPS35 antibody

Excellent
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HeLa)
Specification
HeLa
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.1% Triton X-100
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 22°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Dec 20 2012

Western blot abreview for Anti-VPS35 antibody

Good
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (HeLa)
Loading amount
25 µg
Specification
HeLa
Gel Running Conditions
Reduced Denaturing (12% Tris-Hepes-SDS gel)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Dec 18 2012

Question

will test ab10099 in mouse

Read More

Abcam community

Verified customer

Asked on Oct 05 2011

Answer

Thank you for your phone call. DISCOUNT CODE: xxxx Expiration date: xxx I am very pleased to hear you would like to accept our offer and test ab10099 in mouse. This code will give you 1 free PRIMARY ANTIBODY before the expiration date. To redeem this offer, please submit an Abreview for mouse and include this code in the “Additional Comments” section so we know the Abreview is for this promotion. For more information on how to submit an Abreview, please visit the site: www.abcam.com/Abreviews. Remember, we publish both positive and negative Abreviews on our datasheets so please submit the results of your tests. The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways: 1) Call to place your order and mention the code to our customer service department; 2) Include the code in your fax order; 3) Place your order on the web and enter the promotional code. Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research. The terms and conditions applicable to this offer can be found here: www.abcam.com/collaborationdiscount.

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Answered on Oct 05 2011

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