Confocal immunofluorescent analysis of HeLa cells using Pan-Keratin (C11) Mouse mAb #4545 detected with Anti-Mouse IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 647 Conjugate) (blue). Red = propidium iodide (fluorescent DNA dye).
Flow cytometric analysis of untreated Jurkat cells using Akt (5G3) Mouse mAb #2966 detected with Anti-Mouse IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 647 Conjugate) (green) compared to a nonspecific negative control antibody (red).
High content analysis of C2C12 cells exposed to varying concentrations of U0126 #9903 for 2 hours. With increasing concentrations of U0126, a significant decrease (~20 fold) in phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) signal as compared to the untreated control was observed. When using Phospho-p44/42 MAPK (Erk1/2) protein as a measurement, the IC50 of this compound was 2.5 μM. Data was generated on the Acumen® HCS platform using Anti-Mouse IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 647 Conjugate).
Anti-Mouse IgG (H+L) F(ab')2 Fragment was conjugated to Alexa Fluor® 647 fluorescent dye under optimal conditions and formulated at 2 mg/ml. This F(ab')2 fragment product results in less non-specific binding, as it lacks the Fc domain that can bind to the cells with Fc receptors.
The optimal dilution of the anti-species antibody should be determined for each primary antibody by titration. However, a final dilution of 1:500 – 1:2000 should yield acceptable results for immunofluorescent and flow cytometry assays.
Supplied in 0.1 M sodium phosphate, 0.1 M sodium chloride, pH 7.5, 5 mM sodium azide. Store at 4°C. Do not aliquot the antibody. Protect from light. Do not freeze.
F(ab')2 fragments are prepared from goat antibodies that have been adsorbed against human IgG and human serum.
This product has been optimized for use as a secondary antibody in immunofluorescent applications. Fluorescent anti-species IgG conjugates are ideal for flow cytometry and immunofluorescence. Cell Signaling Technology’s strict quality control procedures assure that each conjugate provides optimal specificity and fluorescence.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
Acumen is a registered trademarks of TTP Labtech.
Alexa Fluor is a registered trademark of Life Technologies Corporation.
The Alexa Fluor dye antibody conjugates in this product are sold under license from Life Technologies Corporation, for research use only for immunocytometry, immunohistochemistry, high content screening (HCS) analysis, or flow cytometry applications.
The transfer of this product is contingent on the buyer using the purchased product solely in research conducted by the buyer (whether the buyer is an academic or for-profit entity), for Immunocytochemistry, high content screening (HCS) analysis, or flow cytometry applications. The sale of this product is expressly conditioned on the buyer not using the product or its components (1) in manufacturing; (2) to provide a service, information, or data to an unaffiliated third party for payment; (3) for therapeutic, diagnostic or prophylactic purposes; (4) resale, whether or not such product or its components are resold for use in research; or for any other commercial purpose. For information on purchasing a license to this product for purposes other than research, contact Life Technologies Corporation, 5791 Van Allen Way, Carlsbad, CA 92008 USA or firstname.lastname@example.org.