I am currently doing a cell adhesion assay with gelatin coating. Can anyone suggest to me a detailed protocol. I have made 1% gelatin by dissolving 1g gelatin in double distilled water (insoluble at this stage) and autoclaved it 121 for 15 mins.The gelatin solution was liquid when I cooled and stored it at 4 degree it was still liquid. I have read it is suppose to solidify is it so? Is it also filtered?
In cell culture normally concentration of gelatin is 0.1% for coating, not 1.0%. I only make the solution and the sterile filtering, no autoclaving step.
Hello Saba, I am using 0.1% gelatin for brain endothelial cells. It worked well, and the well was 80% confluence after 24 hours (1000000 cells in each well/96-well plate). you may use gelatin for most of the adherent cells, but for brain astrocytes I found Fibronictin is the best in teams of growth and viability.
Good luck with your project.
thank you for all the answers. our lab has this protocol which says to use 1% gelatin for coating and for making it I always came across autoclaving but some how everyone suggested me to filter and there was the confusion. Does it look like a white gel after you filter it and store at 4 degree?
thanks peter yes it is milky solution and filtering is a problem :(....I am using 0.45 micron filter to filter it.
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