I don't have an assay for any one particular protein, I just want to western blot all proteins secreted by a tissue sample. If this is feasible, can you give me a protocol or product link?
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judy miller
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Posted 04 June 2004 - 12:11 PM
I'm a lowly undergrad. Please help me out . I am looking to isolate secreted proteins from smooth muscle tissue I don't have an assay for any one particular protein, I just want to western blot all proteins secreted by a tissue sample. If this is feasible, can you give me a protocol or product link?
I don't have an assay for any one particular protein, I just want to western blot all proteins secreted by a tissue sample. If this is feasible, can you give me a protocol or product link?
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phdconsult
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Posted 04 June 2004 - 12:40 PM
Dear Lowly undergrad
What is the primary state of your tissue sample? Frozen/freshly isolated/cell culture/ etc-
What is the primary state of your tissue sample? Frozen/freshly isolated/cell culture/ etc-
Edited by phdconsult, 07 June 2004 - 06:10 PM.
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judy miller
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Posted 07 June 2004 - 04:53 AM
freshly isolated from mice. ( my PI does not want to do a genetic screen - which would make my job a lot easier) . any advice?
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phdconsult
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Posted 07 June 2004 - 04:27 PM
Judy
The first step would be to keep the dynamic action within the tissue going on for some time. You can use Ringer's solution but standard PBS at 37C would work just fine. Allow the freshly isolated tissue to soak in the buffer for 90-120 min at 37C. Recover buffer. Wash the tissue carefully to recover adhering proteins. Proceed to concentrating the solution
Goodluck
The first step would be to keep the dynamic action within the tissue going on for some time. You can use Ringer's solution but standard PBS at 37C would work just fine. Allow the freshly isolated tissue to soak in the buffer for 90-120 min at 37C. Recover buffer. Wash the tissue carefully to recover adhering proteins. Proceed to concentrating the solution
Goodluck
