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The flow cytometry image on the datasheet for ab76409 indicates ab73232 in the text. Is there any data from primary cells in flow cytometry?
Asked on Aug 04 2010
Thank you once again for your recent telephone enquiry. I am pleased to forward the testing flow cytometry protocol for ab76409 IRF3 antibody [EP2419Y] from the originator. Please note, this is a general testing protocol and may require some optimization in your own laboratories. Solutions and reagents 1. 1X PBS buffer 2. Blocking buffer: 0.5% BSA in 1X PBS 3. 2% paraformaldehyde (1% solution – optional for storing samples) 4. 1X FACS permeabilizing solution 5. Fluorescently conjugated secondary antibody Protocol 1. Collect 1 X 106 cells/sample 2. Wash cells once with blocking buffer 3. Fix cells with 2% paraformaldehyde and incubate at room temperature for 10 min 4. Wash cells once with blocking buffer 5. Add 0.5ml 1X FACS permeabilizing solution and incubate at room temperature for 10 min 6. Wash cells once with blocking buffer 7. Incubate cells in blocking buffer for 30 min at room temperature 8. Add primary antibody at the appropriate dilution and incubate for 30 min at room temperature 9. Wash twice with blocking buffer and incubate with fluorescently conjugated secondary antibody for 20 min at room temperature 10. Wash cells twice with blocking buffer 11. Re-suspend cells in 1X PBS and analyze on flow cytometer. Samples can be kept in 1% paraformaldehyde at 4oC overnight I hope this will be helpful. Should you have any further questions, please do not hesitate to contact us.
Answered on Aug 04 2010