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How to extract protein from Perfused brain for western blot?
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+ Western blot
+ Biochemistry
+ Proteins
+ Sequencing
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Kiva
How to extract protein from Perfused brain for western blot?
You can use any cell lysis buffer with protease and phosphatase inhibitors ( Lysis buffer D,CS, RIPA etc.,). It also depends on which protein you are looking for membrane/cytoskeletal/cytoplasmic ?
You can use any cell lysis buffer with protease and phosphatase inhibitors ( Lysis buffer D,CS, RIPA etc.,). It also depends on which protein you are looking for membrane/cytoskeletal/cytoplasmic ?
DearAshwini I am agree with Sebastian If your tissue fixed with formalin,you can not use them. If not, these tissue can be easily use for homogenization and protein extraction. You must lysed the cells using urea and thiourea and concurrently use of protease inhibitor or work on ice under liquid nitrogen.
DearAshwini I am agree with Sebastian If your tissue fixed with formalin,you can not use them. If not, these tissue can be easily use for homogenization and protein extraction. You must lysed the cells using urea and thiourea and concurrently use of protease inhibitor or work on ice under liquid nitrogen.
Maybe you can employ deep sequencing still, but not WB. Hope you good luck. See: Nature. 2014 December 18; 516(7531): 428– 431. doi:10.1038/nature13906.
Maybe you can employ deep sequencing still, but not WB. Hope you good luck. See: Nature. 2014 December 18; 516(7531): 428– 431. doi:10.1038/nature13906.
If you performed the perfusion with (para)formaldehyde, the proteins in your brain sample are chemically crosslinked. I don't think it makes sense to use such a sample for Western Blot.
If you performed the perfusion with (para)formaldehyde, the proteins in your brain sample are chemically crosslinked. I don't think it makes sense to use such a sample for Western Blot.
You can use any cell lysis buffer with protease and phosphatase inhibitors ( Lysis buffer D,CS, RIPA etc.,). It also depends on which protein you are looking for membrane/cytoskeletal/cytoplasmic ?
You can use any cell lysis buffer with protease and phosphatase inhibitors ( Lysis buffer D,CS, RIPA etc.,). It also depends on which protein you are looking for membrane/cytoskeletal/cytoplasmic ?
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DearAshwini
I am agree with Sebastian
If your tissue fixed with formalin,you can not use them.
If not, these tissue can be easily use for homogenization and protein extraction. You must lysed the cells using urea and thiourea and concurrently use of protease inhibitor or work on ice under liquid nitrogen.
DearAshwini
I am agree with Sebastian
If your tissue fixed with formalin,you can not use them.
If not, these tissue can be easily use for homogenization and protein extraction. You must lysed the cells using urea and thiourea and concurrently use of protease inhibitor or work on ice under liquid nitrogen.
More
VOTE
Maybe you can employ deep sequencing still, but not WB. Hope you good luck.
See: Nature. 2014 December 18; 516(7531): 428– 431. doi:10.1038/nature13906.
Maybe you can employ deep sequencing still, but not WB. Hope you good luck.
See: Nature. 2014 December 18; 516(7531): 428– 431. doi:10.1038/nature13906.
More
VOTE
If you performed the perfusion with (para)formaldehyde, the proteins in your brain sample are chemically crosslinked. I don't think it makes sense to use such a sample for Western Blot.
If you performed the perfusion with (para)formaldehyde, the proteins in your brain sample are chemically crosslinked. I don't think it makes sense to use such a sample for Western Blot.
More
VOTE