Secondary Antibodies
Anti-mouse IgG (H+L) (DyLight® 800 Conjugate) #5257
Specificity / Sensitivity
Anti-mouse IgG (H+L) (DyLight® 800 Conjugate) reacts with the heavy and light chains 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 upper panel while corresponding fluorescent western blots are shown in the lower panel.
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 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® 800 Conjugate). DRAQ5® #4084 (fluorescent DNA dye - red) is used for normalization.
Description
Anti-mouse IgG (H+L) was conjugated to DyLight® 800 fluorescent dye under optimal conditions and formulated at 1 mg/ml. Excitation is 777 nm and peak fluorescence emission is 794 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: 1:2000
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
- 4084 DRAQ5®
- 4408 Anti-mouse IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate)
- 4409 Anti-mouse IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 555 Conjugate)
- 4410 Anti-mouse IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 647 Conjugate)
- 4414 Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 647 Conjugate)
- 4416 Anti-rat IgG (H+L), (Alexa Fluor® 488 Conjugate)
- 4417 Anti-rat IgG (H+L), (Alexa Fluor® 555 Conjugate)
- 4418 Anti-rat IgG (H+L), (Alexa Fluor® 647 Conjugate)
- 5151 Anti-rabbit IgG (H+L) (DyLight® 800 Conjugate)
- 5366 Anti-rabbit IgG (H+L) (DyLight® 680 Conjugate)
- 5470 Anti-mouse IgG (H+L) (DyLight® 680 Conjugate)
This product is intended for research purposes only. The product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.
