Confocal immunofluorescent analysis of P19 cells using LIN28A (A177) Antibody #3978 detected with Anti-Rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) (green). Actin filaments have been labeled with DY-554 phalloidin (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) Rabbit mAb #4060 detected with Anti-Rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) and compared to a nonspecific negative control antibody (red).
High content analysis of HeLa cells exposed to varying concentrations of staurosporine for 3hr. With increasing concentrations of staurosporine, a significant decrease (~2.5 fold) in phospho-MAPKAPK-2 signal as compared to the untreated control was observed. When using phospho-MAPKAPK-2 as a measurement, the IC50 of this compound was 92.5 mM. Data were generated on the Acumen HCS platform using Anti-Rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate).
Anti-Rabbit IgG (H+L) F(ab')2 Fragment was conjugated to Alexa Fluor® 488 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.