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Synthetic peptide: SPESEGPIYEGLIL, corresponding to C terminal amino acids 783-796 of Human VPS35.
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.
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.
|IHC-P||Use at an assay dependent concentration. In paraffin embedded Human Testis, shows positive staining in Leydig cells, and cytoplasmic staining (dot-like) of some epithelial cells of the epididymis and of the rete testis.|
|WB||Use a concentration of 0.01 - 1 µg/ml. Detects a band of approximately 85 kDa (predicted molecular weight: 91 kDa).Can be blocked with Human VPS35 peptide (ab23181). Use at a concentration of 1 - 3 µg/ml. Detects a band of approximately 85 kDa observed in human brain and testis lysates (predicted molecular weight: 91 kDa according to NP_060676).|
|ICC/IF||Use at an assay dependent concentration.|
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 goat anti-rabbit IgG (H + L) and goat anti-mouse IgG (H + L) secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging.
ab10099 staining (1
ab10099 staining (1
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