I am trying to perform in vivo plug assay for studying the antiangiogenic effects of my drug. I am using BME culturex (an alternative to Martigel) for the study. I find that after injecting, the gel doesn't solidify at all. Even the bulge disappears after a few hours and I don't see any plug formation. Does anyone have any suggestions? Is my matrigel degraded?
Hello Abu Sufiyan Chhipa
One reason I can think of could perhaps be the protein concentration of BME cultrex that you are using. Every lot has a specific protein concentration or dilution factor included on the Certificate of Analysis. The protein concentration varies between lots and this specific protein concentration should be used to calculate the amount of BME cultrex required for your application. The optimal protein concentration is application dependent.
To prevent incomplete gel formation for in vivo applications, Matrigel matrix should not be diluted to a final concentration below 4 mg/ml. For BME Cultrex, the final concentration should be around 7-10mg/ml. Determine the protein concentration range that works best for your application. To prevent incomplete gel formation for in vivo applications, do not dilute by fold dilution but dilute to a specific concentration (mg/ml).
Hope this information will be helpful!
Best.
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