Anti-VPS26 抗体 (ab23892)

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) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

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

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).

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.

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Immunocytochemical analysis of Rat NRK cell line, labeling VPS26 with ab23892. Cells were formaldehyde fixed, permeabilized with 0.1% Triton X-100 in PBS for 3 minutes and blocked with 1% BSA for 30 minutes at 25°C. Cells were incubated with ab23892 diluted 1/250 in 1% BSA in PBS for 1 hour at 25°C.

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