The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
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.
1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Use a concentration of 1 µg/ml.
Use at 1 µg/mg of lysate.
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).
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
VPS 26 antibody
VPS 26A antibody
Anti-VPS26 antibody images
Western blot - Anti-VPS26 antibody (ab23892)
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) Lanes 1 - 3: Merged signal (red and green). Green - ab23892 observed at 40 kDa. Red - loading control, ab8245, observed at 37 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 ab8245 (loading control to GAPDH) 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.
Western blot - VPS26 antibody (ab23892)
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
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).
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
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.
Western blot - Anti-VPS26 antibody (ab23892)This image is courtesy of an anonymous Abreview
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
Immunocytochemistry/ Immunofluorescence - Anti-VPS26 antibody (ab23892)Image from Xia B et al., PLoS One. 2010 Jul 23;5(7):e11771. Fig 3.; doi:10.1371/journal.pone.0011771; July 23, 2010, PLoS ONE 5(7): e11771.
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 Chem291:18283-98 (2016).
Read more (PubMed: 27385586) »