Mycophenolic Acid is an antibiotic useful in research for the selection of animal cells that express the E. coli gene coding for XGPRT (xanthine guanine phosphoribosyltransferase). Mycophenolic Acid is also used as immunosuppressive agent becasue it acts by inhibiting lymphocyte proliferation and antibody production. Cells transfected with the E. coli GPT gene can be selected for growth in medium containing GPT Selection Reagent. Each Kit Contains:• 500X Mycophenolic Acid containing ETOH (12.5mg/mL) 2 x 1mL vials• 1 bottle (10mL) of 100X Aminopterin solution which consists of: - xanthine 25 mg/mL - hypoxanthine: 1.5 mg/mL - aminopterin: 0.2mg/mL - thymidine: 1 mg/mL - in 0.08M NaOH
Synonyms: MPA & Aminopterin solution
Application: 1. Seed approximately 106 cells on 100 mm culture plates in Dulbecco's modified Eagle medium (DMEM) containing 5% fetal calf serum and penicillin/streptomycin, and allow them to grow for twenty-four hours. 2. Transfect the culture with 10-20 mug of the plasmid-EcoGPT DNA. 3. After 3 days at 37C in DMEM containing 5% fetal calf serum, trypsinize the cell monolayers and disperse 5 x 105 cells on fresh 100 mm culture plates in DMEM containing 10% dialyzed fetal calf serum supplemented with the MPA and Aminopterin solutions. This will give a final solution of approximately: 250 mug/mL xanthine, 15 mug/mL hypoxantine, 10 mug/mL thymidine, 2 mug/mL aminopterin, and 25 mug/mL mycophenolic acid. Dialyzed serum is used to speed the selection process. Selection will be slower with undialyzed serum. 4. Twenty-four hours later, replace the medium with fresh medium containing the same supplements, and fluid change. 5. Repeat every 3 days. Colonies are visible in 7-10 days, typically. Note: Other media bases can also be used such as Ham's F12. Much depends upon the cell type being used.