ISISBio:Protocols/p11 resin preparation

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Contents

Overview

Preparation of Whatmann P11 Phosphocellulose resin for cation exchange chromatography.

Phosphocellulose remains a useful and cheap cation exchange resin and is particularly good for the final stages of purification of DNA binding proteins. The preparation of the resin for optimum performance is somewhat tedious but carefull attention to detail will yield a higher performance coplumn with lower back pressure.

Materials

  • Whatmann P11 Phosphocelluose resin
  • Purified water
  • 0.2 M NaOH
  • 0.2 M HCl
  • Storage buffer (Tris-HCl pH 7.6 is a common choice)

Procedure

  1. Weigh out approximately as many grams of resin as millilitres of column will be required into a large container with at least 20-fold larger volume
  2. Suspend the resin in at least 20 volumes of water and allow to settle for at least 30 minutes
    1. Pour off supernatant, resuspend and repeat at least 8-10 times.
    2. Step 3 has multiple sub-steps within it.
    3. Enumerate each of those.

Notes

  1. List troubleshooting tips here.
  2. You can also link to FAQs/tips provided by other sources such as the manufacturer or other websites.
  3. Anecdotal observations that might be of use to others can also be posted here.

Please sign your name to your note by adding '''*~~~~''': to the beginning of your tip.

References

Relevant papers and books

  1. Goldbeter A and Koshland DE Jr. . pmid:6947258. PubMed HubMed [Goldbeter-PNAS-1981]
  2. JACOB F and MONOD J. . pmid:13718526. PubMed HubMed [Jacob-JMB-1961]
  3. Mark Ptashne. A genetic switch. Cold Spring Harbor, N.Y.: Cold Spring Harbor Laboratory Press, 2004. isbn:0879697164. [Ptashne-Genetic-Switch]
All Medline abstracts: PubMed HubMed

Contact

  • Who has experience with this protocol?

or instead, discuss this protocol.


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