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Synthetic peptide designed within the sequence of human SQSTM1
This product is a recombinant rabbit monoclonal antibody.
Produced using Abcam’s RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5,675,063 and/or 7,429,487.
Alternative versions available:
Anti-SQSTM1 / p62 antibody - Autophagosome Marker (Alexa Fluor® 594) [EPR4844] (ab203429)
Anti-SQSTM1 / p62 antibody (Alexa Fluor® 647) [EPR4844] (ab194721)
Anti-SQSTM1 / p62 antibody (HRP) [EPR4844] (ab194720)
Our Abpromise guarantee covers the use of ab109012 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/10000 - 1/50000. Detects a band of approximately 62 kDa.|
|Flow Cyt||1/10 - 1/100.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
|ICC/IF||1/100 - 1/500.|
Lane 1: Wild type HAP1 whole cell lysate (20 µg)
Lane 2: SQSTM1 knockout HAP1 whole cell lysate (20 µg)
Lane 3: HeLa whole cell lysate (20 µg)
Lane 4: HepG2 whole cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab109012 observed at 47 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab109012 was shown to specifically react with SQSTM1 when SQSTM1 knockout samples were used. Wild-type and SQSTM1 knockout samples were subjected to SDS-PAGE. Ab109012 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 10000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling Autophagosome Marker with ab109012 at 1/500 dilution (4.6μg/ml). Cells were fixed in 100% Methanol. ab150077 an AlexaFluor® 488 Goat anti-Rabbit secondary antibody was used at 1/1000 dilution (2 μg/ml). DAPI was used to counter stain the nucleus. Confocal image showing cytoplasmic staining on HeLa cell line.
Overlay histogram showing HeLa cells stained with ab109012 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab109012, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"