Confocal immunofluorescent analysis of HeLa cells using COX IV (3E11) Rabbit mAb #4850 detected with Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 594 Conjugate) (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Flow cytometric analysis of Jurkat cells, untreated (green) or treated with LY294002 #9901, Wortmannin #9951, and U0126 #9903 (blue), using Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb #4060 detected with Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 594 Conjugate).
Anti-rabbit IgG (H+L), F(ab')2 Fragment was conjugated to Alexa Fluor® 594 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 pooled human serum, mouse serum, plasmacytoma/hybridoma proteins, and purified human paraproteins.
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.