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Please confirm if this has functional activity in cell culture (Functional studies listed as tested application)
Asked on Jul 03 2008
Thank you for your telephone enquiry yesterday afternoon. I can confirm that our ab50049 WISP3 human protein (Active) has been tested for biological activity in cell culture, so it will be covered by our guarantee for this application. The originator of the protein has kindly provided the following information which I hope you will find helpful: The ED50 was determined by the dose-dependent proliferation of the MCF-7 cell line. The expected ED50 for this effect is 0.2-0.3 µg/ml. The protocol used was as follows: Note: As of lot 020405 culture and assay media have been changed to media containing no phenol red. Also the cells are cultured in 10% Certified FBS, but the assay media contains 0.25% HI-FBS. 1. Resuspend cells in assay media containing 0.25% HI-FBS Transfer 100ul culture/well to assay plate (5,000 cells/well; passage 22). 2. Serial dilute h-WISP-3 in assay media in duplicate. 3. Add 100ul/well of h-WISP-3 to cells in assay plate. Final assay volume is 200ul/well containing 0.25% HI-FBS, and h-WISP-3. 4. Incubate 90 hours. 5. Add 20ul of Substrate Cell Titer Aqueous One Solution Reagent to each well. 6. Incubate 37oC and read OC at 490nm. The endotoxin level was tested for this and was found to be less than 0.1 ng per µg (1EU/µg). I hope this information is useful to you. Please do not hesitate to contact us should you have any additional questions.
Answered on Jul 03 2008