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Message | User | Date(yyyy-mm-dd) |
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Collagen Applications??? | rpallela | 2007-12-07 | Click here to register. |
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Hi all, I have purified collagen from invertebrate sources. I prefer to check the adhrence activity of the collagens to any of the cell lines. But is this a valauable parameter for the collagen functional activity? and also please suggest me, how to formulate/dispense the purified collagens to form layer (any protocaol?)in culture flasks? and what are the physical and other factors to be taken care of.....?
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| trish | 2007-12-07 | Click here to register. |
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 | I haven't worked with invertebrate collagens and I am unsure how much of my knowledge about vertebrate molecules will extend to your interests.
Attachment of cells to collagens are not fully dependent on the structure of the collagen. The integrins that are resposible for the attachement recognize specific amino acid sequences and do not require an intact functional triple helical molecule to attach.
The purified collagens are diluted to the concentration that you want in an acidic solution. The plastic plate is flooded with the collagen solution and allowed to sit at room temperature for at least 10 minutes. The plates should be rinsed with PBS or other buffer to remove the acidic residue...I like to use a buffer with a pH indicator to make sure that I have neutralized the plate...and allowed to air dry. Coated plates can be stored at 4C for several weeks. |
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| rpallela | 2007-12-13 | Click here to register. |
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| Hi trish
Thank u for the valuable information. Please provide any references for the information you have provided, which could be useful before i initiate the experiment.
Rpallela |
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| anildsgs | 2007-12-19 | Click here to register. |
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| Hi,
I havent used any collegen based cell proliferation in my cell culutre experiments. I am working on murine & human mesenchymal stem cells, to proliferate and further diferentiate into different lineages. can you suggest me how shall i proceed with collegen based cell cultures. is any commercial avaible collegen is recomended or we can prepare in house. kindly give me details.
thanks in advance.
Anil Kumar
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