Full length native protein (purified) corresponding to Src. Antibody generated by immunizing BALB/c mice with full length native protein (purified) and then fusing with P3X63 Ag8.653 myeloma cells.
May appear as a doublet due to phosphorylation. May be used to precipitate active Src which can then be used in a kinase reaction.
Abcam is committed to meeting high standards of ethical manufacturing and has decided to discontinue this product by June 2019 as it has been generated by the ascites method. We are sorry for any inconvenience this may cause. We would recommend antibody ab222220 as a replacement.
Our Abpromise guarantee covers the use of ab16885 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
アプリケーション | Abreviews | 特記事項 |
---|---|---|
Flow Cyt | Use 1µg for 106 cells. ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody. |
|
WB | Use a concentration of 2.5 µg/ml. Predicted molecular weight: 59.7 kDa. ab16885 may appear as a doublet on WB due to phosphorylation. This antibody can be used to precipitate active src kinase. | |
IP | Use at 5 µg/mg of lysate. See Abreview. |
Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: Src knockout HAP1 cell lysate (20 µg)
Lane 3: HeLa cell lysate (20 µg)
Lane 4: A431 cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab16885 observed at 60 kDa. Red - loading control, ab8227, observed at 42 kDa.
ab16885 was shown to specifically react with Src when Src knockout samples were used. Wild-type and Src knockout samples were subjected to SDS-PAGE. ab16885 and ab8227 (loading control to beta Actin) were diluted 2.5µg/ml and 1/1000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) and Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) secondary antibodies at 1/10000 dilution for 1 h at room temperature before imaging.
Overlay histogram showing SH-SY5Y cells stained with ab16885 (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 (ab16885, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.
Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: Src knockout HAP1 cell lysate (20 µg)
Lane 3: HeLa cell lysate (20 µg)
Lane 4: A431 cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab16885 observed at 60 kDa. Red - loading control, ab8245, observed at 37 kDa.
This western blot image is a comparison between ab16885 and a competitor's top cited rabbit polyclonal antibody.
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