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neutralization assay: 1) what antibody conc. should be used? 2) for a 3-5 day exp. (with human islet cell in suspension) does the antibody need to be added daily or just once during that time?
Asked on Apr 30 2010
Thank you for your inquiry. I have received the following answers to your questions from the lab: 1) The actual concentration has to be determined by the end users due to the variation of cytokine concentration and cell types used. But in human CXCR2 transfected BaF/3 cells, approximately 0.1-0.3 μg/mL of the antibody neutralizes 50% of the bioactivity due to 20 ng/mL of recombinant human CXCL8. 2) The antibody will be still active after 3 days in the culture system, but if CXCL8 is produced by the cells used in your study, it will be appropriate to add extra antibody, but again, this has to be determined by the end users based on their own study. I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.
Answered on Apr 30 2010