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protein extraction from tumor samples containing matrigel?


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2 replies to this topic

#1 Thapa

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Posted 28 May 2009 - 06:15 PM

Hello there,
I have snap-frozen tumor (from HT29 cells injected with matrigel). I wish to process those samples for western blot by extracting total proteins. For this, do I need to remove matrigel from tumor tisssue? Or I can lyse them directly?

If I lyse them prior to removing matrigel, is it hinders latter processes ??

Thank you in advance.

Best regards,
Thapa
"Learning without thought is labor lost"

#2 alan6017518

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Posted 29 May 2009 - 07:22 AM

You injected the cells with Matrigel? I have never heard of that, I thought you can only grow cells on Matrigel, I have never worked with it before. But I did grow HT-29 cells long time ago

May be you want to check what is the protein content of the Matrigel, if it is not containing the protein you are looking for then I guess its fine to treat the sample with the gel in it.

Hope these help.

#3 Agonist

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Posted 03 June 2009 - 05:50 PM

You injected the cells with Matrigel? I have never heard of that, I thought you can only grow cells on Matrigel, I have never worked with it before. But I did grow HT-29 cells long time ago

May be you want to check what is the protein content of the Matrigel, if it is not containing the protein you are looking for then I guess its fine to treat the sample with the gel in it.

Hope these help.


Hey friend alan,
I think Thapa is right in his problem. If I am not wrong, he has implanted tumor cells mixed with growth factor reduced matrigel to study in vivo tumor growth and angiogenesis. Just what he asked here that how to process his excised tumors for western blot analysis?? More in simple, he has tumor tissue containing matrigel and he wants to avoid matrigel before the lysis of tumor cells with appropriate buffer.

Me? truly not experienced such experiment. But I think that just homogenzitation with suitable buffer and lysis would be okay. In general, however, reports say that keeping matrigel overnight at 4C to melt down ....but not sure how much it willbe separted from tumor cells.

I hope Bioforumers doing tumor works will share more informations.

Best luck..
"Learning without thought is labour lost"
http://labofphysiology.blogspot.com/




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