Secondary Antibodies
Anti-mouse IgG (H+L) (DyLight® 680 Conjugate) #5470
Specificity / Sensitivity
Anti-mouse IgG (H+L) (DyLight® 680 Conjugate) reacts with heavy and light chain of most mouse immunoglobulins. No cross-reactivity to other serum proteins has been detected. This antibody may cross-react with immunoglobulins from other species.
Source / Purification
This antibody is prepared from goat antibodies and purified by immunoaffinity chromatography using antigen coupled to agarose beads.
Fluorescent Detection
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 Phospho-p44/42 MAPK (Erk1/2) (Thr202/204) (D13.14.4E) XP® Rabbit mAb #4370 detected with Anti-rabbit IgG (H+L) (DyLight® 800 Conjugate) #5151 (green) and p44/42 MAPK (Erk1/2) (3A7) Mouse mAb #9107 detected with Anti-mouse IgG (H+L) (DyLight® 680 Conjugate) (red). The array image pixel intensities obtained using a LI-COR® Biosciences Odyssey® Infrared Imaging System are shown in the top figure while corresponding fluorescent western blots are shown in the bottom figure.
In-Cell Western
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 (~2.5 fold) in Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (E10) Mouse mAb #9106 signal as compared to the TPA-stimulated control was observed. Data and images were generated on the LI-COR® Biosciences Odyssey® Infrared Imaging System using Anti-mouse IgG (H+L) (DyLight® 680 Conjugate).
Description
Anti-mouse 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.
Directions for Use
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
Applications
This product has been optimized for use as a secondary antibody in fluorescent western blotting and In-Cell Western™.
Background
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.
Application References
Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!
Companion Products
For Research Use Only. Not For Use In Diagnostic Procedures.