Confocal immunofluorescent analysis of HeLa cells labeled with MEK1/2 (47E6) Rabbit mAb #9126 detected with Anti-Rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 555 Conjugate) (red, left) compared to an isotype control (right). Actin filaments have been labeled with fluorescein phalloidin (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Confocal immunofluorescent analysis of mouse cerebellum using α-Synuclein Antibody (IF Preferred) #2628 detected with Anti-Rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 555 Conjugate) (red) and Neurofilament-L (DA2) Mouse mAb #2835 detected with Anti-Mouse IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4408 (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
High content analysis of A549 cells exposed to varying concentrations of LY294002 #9901 for 3 hrs, followed by 100 ng/mL EGF for 20 minutes. With increasing concentrations of LY294002, a significant decrease (~5 fold) in phospho-S6 Ribosomal Protein (Ser235/236) signal as compared to the uninhibited control was observed. When using phospho-S6 as a measurement, the IC50 of this compound was 3.06 μM. Data were generated on the Acumen HCS platform using Anti-Rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 555 Conjugate).
Anti-Rabbit IgG (H+L) F(ab')2 Fragment was conjugated to Alexa Fluor® 555 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 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.