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Recombinant full length protein corresponding to Human Cyclin D3.
MELLCCEGTRHAPRAGPDPRLLGDQRVLQSLLRLEERYVPRASYFQCVQR EIKPHMRKML AYWMLEVCEEQRCEEEVFPLAMNYLDRYLSCVPTRKAQ LQLLGAVCMLLASKLRETTPLT IEKLCIYTDHAVSPRQLRDWEVLVLG KLKWDLAAVIAHDFLAFILHRLSLPRDRQALVKK HAQTFLALCATDYT FAMYPPSMIATGSIGAAVQGLGACSMSGDELTELLAGITGTEVDCL RA CQEQIEAALRESLREASQTSSSPAPKAPRGSSSQGPSQTSTPTDVTAIHL
Our Abpromise guarantee covers the use of ab28283 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|Flow Cyt||Use 1µg for 106 cells. ab170190-Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.|
|WB||Use at an assay dependent concentration. Predicted molecular weight: 35 kDa.|
|IP||Use at an assay dependent concentration.|
|IHC-P||Use at an assay dependent concentration.|
Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
Lane 2: Cyclin D3 (KO) knockout HAP1 whole cell lysate (20 µg)
Lane 3: HEK293 whole cell lysate (20 µg)
Lane 4: Jurkat whole cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab28283 observed at 33 kDa. Red - loading control, ab176560, observed at 50 kDa.
ab28283 was shown to specifically recognize CCND3 in wild-type HAP1 cells along with additional cross reactive bands. No bands was observed when CCND3 knockout samples were uexamined. Wild-type and CCND3 knockout samples were subjected to SDS-PAGE. Ab28283 and ab176560 (Rabbit anti alpha Tubulin loading control) were incubated overnight at 4°C at 1 ug/ml and 1/10,000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed ab216772 and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed ab216777 secondary antibodies at 1/20,000 dilution for 1 hour at room temperature before imaging.
Overlay histogram showing HeLa cells stained with ab28283 (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 (ab28283, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was a goat anti-mouse DyLight® 488 (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.
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