Confocal immunofluorescent analysis of mouse pons using CaMKII-α (6G9) Mouse mAb #50049 detected with Anti-mouse IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 555 Conjugate) #4409 (red) and GFAP (D1F4Q) XP® Rabbit mAb #12389 detected with Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 647 Conjugate) #4410 (green pseudocolor). Sections were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
Confocal immunofluorescent analysis of HeLa cells using S6 Ribosomal Protein (54D2) Mouse mAb #2317 detected with Anti-Mouse 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. Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
High content analysis of C2C12 cells exposed to varying concentrations of LY294002 #9901 for 2 hours. With increasing concentrations of LY294002, a significant decrease (~20 fold) in phospho-S6 Ribosomal protein (Ser235/236) signal as compared to the untreated control was observed. When using phospho-S6 Ribosomal protein as a measurement, the IC50 of this compound was 2.3 μM. Data was generated on the Acumen® HCS platform using Anti-Mouse IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 555 Conjugate).
Anti-Mouse 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 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.
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