Overview

Properties

Applications

Our Abpromise guarantee covers the use of ab23892 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
WB Use a concentration of 1 µg/ml. Detects a band of approximately 40 kDa (predicted molecular weight: 40 kDa).

Abcam recommends using milk as the blocking agent.

IHC-P 1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ICC/IF Use a concentration of 1 µg/ml.
IP Use at 1 µg/mg of lysate.

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).
  • Sequence similaritiesBelongs to the VPS26 family.
  • Cellular localizationCytoplasm. Endosome membrane.
  • Information by UniProt
  • Database links
  • Alternative names
    • FLJ12930 antibody
    • HB 58 antibody
    • HB58 antibody
    • Hbeta 58 antibody
    • Hbeta 58 homolog antibody
    • Hbeta58 antibody
    • Hbeta58 homolog antibody
    • hVPS 26 antibody
    • hVPS26 antibody
    • MGC94168 antibody
    • PEP 8A antibody
    • PEP8A antibody
    • Vacuolar protein sorting 26 A antibody
    • Vacuolar protein sorting 26 antibody
    • Vacuolar protein sorting 26 homolog A antibody
    • Vacuolar protein sorting 26 homolog A S. pombe antibody
    • Vacuolar protein sorting 26 homolog A yeast antibody
    • Vacuolar protein sorting 26 yeast antibody
    • Vacuolar protein sorting 26 yeast homolog antibody
    • Vacuolar protein sorting 26A antibody
    • Vacuolar protein sorting associated protein 26A antibody
    • Vacuolar protein sorting-associated protein 26A antibody
    • Vesicle protein sorting 26 antibody
    • Vesicle protein sorting 26A antibody
    • VP26A_HUMAN antibody
    • VPS 26 antibody
    • VPS 26A antibody
    • VPS26A antibody
    see all

Anti-VPS26 antibody images



  • Predicted band size : 40 kDa

    Lane 1: Wild-type HAP1 cell lysate (20 µg)
    Lane 2: VPS26 knockout HAP1 cell lysate (20 µg)
    Lane 3: Jurkat cell lysate (20 µg)
    Lane 4: K562 cell lysate (20 µg)
    Lanes 1 - 4: Merged signal (red and green). Green - ab23892 observed at 124 kDa. Red - loading control, ab18058, observed at 40 kDa.

    ab23892 was shown to recognize VPS26 when VPS26 knockout samples were used, along with additional cross-reactive bands. Wild-type and VPS26 knockout samples were subjected to SDS-PAGE. ab23892 at a concentration of 1 µg/ml  and ab18058 (loading control to Vinculin) diluted to 1/10000 were incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

  • All lanes : Anti-VPS26 antibody (ab23892) at 1 µg/ml

    Lane 1 : Human kidney lysate
    Lane 2 : Human kidney lysate with Human VPS26 peptide (ab24288) at 1 µg/ml

    Lysates/proteins at 20 µg per lane.

    Secondary
    Alexa Fluor Goat polyclonal to Rabbit IgG (700) at 1/10000 dilution

    Performed under reducing conditions.

    Predicted band size : 40 kDa
    Observed band size : 37 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 25 kDa,50 kDa. We are unsure as to the identity of these extra bands.

    ab23892 detects a band of ~ 38kDa in Human kidney lysate. This band is quenched by the addition of the immunizing peptide, ab24288. ab23892 also detects a band of the correct size in HEK293 and mouse kidney lysate (data not shown).

    See Abreview

  • Image courtesy of Human Protein Atlas

    ab23892 staining VPS26 in Human kidney. The paraffin embedded tissue was incubated with ab23892 (1/500 dilution) for 30 mins at room temperature. Antigen retrieval was performed by heat induction in citrate buffer pH 6. ab23892 was tested in a tissue microarray (TMA) containing a wide range of normal and cancer tissues as well as a cell microarray consisting of a range of commonly used, well characterised human cell lines

    Further results for this antibody can be found at www.proteinatlas.org

  • ICC/IF image of ab23892 stained human HEK 293 cells. The cells were 4% PFA fixed (10 min), permabilised in 0.1% PBS-Tween (20 min) and incubated with the antibody (ab23892, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue). This antibody also gave a positive IF result in HeLa and HepG2 cells.

  • All lanes : Anti-VPS26 antibody (ab23892) at 1/1000 dilution

    Lane 1 : Rat NRK whole cell lysate at 25 µg
    Lane 2 : Rat NRK whole cell lysate at 50 µg

    Secondary
    HRP-conjugated goat anti-rabbit polyclonal IgG at 1/5000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 40 kDa
    Observed band size : 40 kDa


    Exposure time : 10 seconds

    This image is courtesy of an anonymous Abreview

    Blocked with 5% milk for 30 min at 25°C

    See Abreview

  • All lanes : Anti-VPS26 antibody (ab23892) at 1/1000 dilution

    Lane 1 : HEK293T whole cell lysate
    Lane 2 : HeLa whole cell lysate

    Lysates/proteins at 50 µg per lane.

    Secondary
    HRP-conjugated goat anti-rabbit polyclonal IgG at 1/5000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 40 kDa
    Observed band size : 40 kDa


    Exposure time : 2 seconds

    This image is courtesy of an anonymous Abreview

    Blocked with 5% milk for 30 min at 25°C

    See Abreview

  • Immunofluorescence analysis of HeLa cells treated with mDpy-30 siRNA, staining VPS26 (green) with ab23892.

    Cells were fixed formaldehyde, permeabilized and blocked with 5% goat serum for 20 minutes. Samples were incubated with primary antibody in blocking buffer for 2 hours before incubating with fluorophore-conjugated secondary antibody for 1 hour. Nuclei were detected using DAPI.

References for Anti-VPS26 antibody (ab23892)

This product has been referenced in:
  • Follett J  et al. Parkinson Disease-linked Vps35 R524W Mutation Impairs the Endosomal Association of Retromer and Induces a-Synuclein Aggregation. J Biol Chem 291:18283-98 (2016). Read more (PubMed: 27385586) »
  • Da Ros M  et al. Retromer vesicles interact with RNA granules in haploid male germ cells. Mol Cell Endocrinol 401:73-83 (2015). Read more (PubMed: 25486514) »

See all 10 Publications for this product

Product Wall

Application Immunoprecipitation
Sample Human Cell lysate - whole cell (CHRF)
Total protein in input 500 µg
Immuno-precipitation step Protein A/G
Specification CHRF
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Submitted Sep 14 2015

Application Immunocytochemistry/ Immunofluorescence
Sample African Green Monkey Cell (COS)
Specification COS
Fixative Formaldehyde
Permeabilization Yes - 0.1% Triton X-100 in PBS for 3 min
Blocking step BSA as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: 25°C
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Submitted Sep 28 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (HeLa)
Specification HeLa
Fixative Formaldehyde
Permeabilization Yes - 0.1% Triton X-100 in PBS for 3 min
Blocking step BSA as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: 25°C
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Submitted Sep 26 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Mouse Cell (3T3)
Specification 3T3
Fixative Formaldehyde
Permeabilization Yes - 0.1% Triton X-100 in PBS for 3 min
Blocking step BSA as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: 25°C
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Submitted Sep 26 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Rat Cell (NRK)
Specification NRK
Fixative Formaldehyde
Permeabilization Yes - 0.1% Triton X-100 in PBS for 3 min
Blocking step BSA as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: 25°C
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Submitted Sep 26 2012

Application Immunocytochemistry/ Immunofluorescence
Sample Chinese Hamster Cell (CHO)
Specification CHO
Fixative Formaldehyde
Permeabilization Yes - 0.1% Triton X-100 in PBS for 3 min
Blocking step BSA as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: 25°C
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Submitted Sep 26 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (HEK293F and HeLa)
Loading amount 50 µg
Specification HEK293F and HeLa
Gel Running Conditions Reduced Denaturing (10% Tris-glycine gel)
Blocking step Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
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Submitted Sep 17 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Mouse Cell lysate - whole cell (3T3)
Loading amount 75 µg
Specification 3T3
Gel Running Conditions Reduced Denaturing (10% Tris-glycine gel)
Blocking step Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
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Submitted Sep 17 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Rat Cell lysate - whole cell (NRK)
Loading amount 25 µg
Specification NRK
Gel Running Conditions Reduced Denaturing (10% Tris-glycine gel)
Blocking step Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
Username

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Submitted Sep 17 2012

Application Western blot
Sample African Green Monkey Cell lysate - whole cell (COS)
Loading amount 50 µg
Specification COS
Gel Running Conditions Reduced Denaturing
Blocking step Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
Username

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Submitted Sep 17 2012

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"