Western blot analysis of Jurkat cell lysates (#9194) treated with either U0126 (MEK 1/2 inhibitor) #9903 or TPA (12-O-Tetradecanoylphorbol-13-Acetate) #4174 using p44/42 MAPK (Erk1/2) (137F5) Rabbit mAb #4695 detected with Anti-rabbit IgG (H+L) (DyLight™ 680 Conjugate) (red) and Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (E10) Mouse mAb #9106 detected with Anti-mouse IgG (H+L) (DyLight™ 800 Conjugate) #5257 (green). The array image pixel intensities obtained using a LI-COR® Biosciences Odyssey® Infrared Imaging System are shown in the upper panel while corresponding fluorescent western blots are shown in the lower panel.Learn more about how we get our images
In-Cell Western™ analysis of A549 cells exposed to varying concentrations of U0126 (MEK1/2 Inhibitor) #9903 for 3 hours, followed by TPA (Phorbol-12-Myristate-13-Acetate) #9905 stimulation for 30 minutes. With increasing concentrations of U0126, a significant decrease (~5 fold) in Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb #4370 signal as compared to the TPA-stimulated control was observed. When using phospho-Erk as a measurement, the IC50 of this compound was 1.9 μM. Data and images were generated on the LI-COR® Biosciences Odyssey® Infrared Imaging System using Anti-rabbit IgG (H+L) (DyLight™ 680 Conjugate).Learn more about how we get our images
The optimal dilution of the anti-species antibody should be determined by the user. However, the final dilutions below should yield acceptable results for the respective applications.
Fluorescent western blotting: 1:15000
In-Cell Western: 1:500
Supplied in 100 mM PBS, pH 7.2, containing 1% BSA and 0.02% sodium azide. Store at 4°C. Protect from light. Do not freeze.
Anti-rabbit IgG (H+L) was conjugated to DyLight™ 680 fluorescent dye under optimal conditions and formulated at 1 mg/ml. Excitation is 684 nm and peak fluorescence emission is 715 nm.
Anti-rabbit IgG (H+L) (DyLight™ 680 Conjugate) reacts with heavy and light chain of most rabbit immunoglobulins. No cross-reactivity to other serum proteins has been detected. This antibody may cross-react with immunoglobulins from other species.
This antibody is prepared from goat antibodies and purified by immunoaffinity chromatography using antigen coupled to agarose beads.
Near infrared anti-species IgG conjugates are ideal for fluorescent western blotting and In-Cell Western. Cell Signaling Technology's strict quality control procedures assure that each conjugate provides optimal specificity and fluorescence.
This product has been optimized for use as a secondary antibody in fluorescent western blotting and In-Cell Western™.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc. DyLight is a trademark of Thermo Fisher Scientific, Inc. and its subsidiaries. LI-COR is a registered trademark of LI-COR, Inc. Odyssey is a registered trademark of LI-COR, Inc.
Discover what’s going on at CST, receive our latest application notes and tips, read our science features, and learn about our products.
To get local purchase information on this product, click here.