Sample Diluent Optimization Pack

1. Prepare samples and controls., 2. Dilute 10X Apoptosis Wash Buffer 1:10 with diH20., 3. Reconstitute FLICA with 50 μL DMSO., 4. Dilute FLICA 1:5 by adding 200 μL PBS., 5. Add diluted FLICA to each sample at 1:30 – 1:60. For example, to stain at 1:30, add 10 μL to 290 μL of cultured cells. To stain at 1:60, add 5 μL to 295 μL of cultured cells., 6. Incubate approximately 1 hour., 7. Remove media and wash cells 3 times: add 1X Apoptosis Wash Buff er and spin cells., 8. If desired, label with additional stains, such as Hoechst, 7-AAD, or an antibody., 9. If desired, fix or embed cells., 10. Analyze with a fluorescence microscope, fluorescence plate reader, or flow cytometer. SR-FLICA excites at 550-580 nm and emits at 590-600 nm.

When developing any new ELISA, it is often helpful to compare a number of different sample diluent formulations to better match the matrix complexity present in the sample Wells with the standard curve calibrator matrix Wells. Most of the time invested in developing a new ELISA/EIA/RIA will involve selecting the proper calibrator matrix that matches most closely to the often very complex biological sample matrix that is being evaluated. Comparing three different sample/calibrator diluent formulations on the same IgG or antigen-coated ELISA plate makes it is easy to determine which diluent will best equalize the calibrator Well antibody-binding kinetics with the biological sample/ dilution Well antibody-binding kinetics. The Sample Diluent Optimization Pack provides 100 mL of each of our three Sample Diluent formulations to begin optimizing the sample diluent portion of the assay development process.