Team:Berkeley Wetlab/Passenger: TypeIII Needle scFv

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(Difference between revisions)
(New page: ===dot blot and visualization=== #pipet 5ul of cell lysate onto nitrocellulose paper and allow to dry completely<br> #wet nitrocellulose with 10ml of 1X TBST for 5minutes<br> #block with 2...)
(→dot blot and visualization)
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===incubation with espA protein===
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#
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===dot blot and visualization===
===dot blot and visualization===
#pipet 5ul of cell lysate onto nitrocellulose paper and allow to dry completely<br>
#pipet 5ul of cell lysate onto nitrocellulose paper and allow to dry completely<br>
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#leave the last wash on for developing
#leave the last wash on for developing
#add 3mls of chemiluminescent reagents to the nitrocellulose paper and take pictures
#add 3mls of chemiluminescent reagents to the nitrocellulose paper and take pictures
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 +
[TBST recipe]

Revision as of 10:10, 13 October 2009

incubation with espA protein

dot blot and visualization

  1. pipet 5ul of cell lysate onto nitrocellulose paper and allow to dry completely
  2. wet nitrocellulose with 10ml of 1X TBST for 5minutes
  3. block with 2% BSA (.2g BSA in 10ml 1X TBST)for 1 hour
  4. wash 3X with TBST - 5minutes/wash
  5. add 4ml of 1:500 Anti-His HRP: TBST, wait 30minutes
  6. wash 3X with TBST - 5minutes/wash
  7. leave the last wash on for developing
  8. add 3mls of chemiluminescent reagents to the nitrocellulose paper and take pictures

[TBST recipe]

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