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Anti-Ubiquitin antibody (ab19247)

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This product is covered by the Abpromise guarantee. Our scientific support team are available to answer any questions or queries - fill out an inquiry form for ab19247 for help.

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2 questions for ab19247

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Question 1

Wednesday 20-December-2006

Conditions employed for WB testing. Customer is using S.cerevisiae

ANSWER:

  

Thank you for your enquiry.

Further to correspondence with the source of this antibody I can tell you that the following approach was employed for the testing of this antibody by western blot analysis. In brief the samples were prepared in denaturing (and what I am assuming, reduced conditions and electrophoresed under denaturing conditions;

1. Cells were washed twice by centrifugation with PBS. Total cell extracts were prepared by suspending cell pellets in RIPA buffer (50mM Tris, pH 7.2, 150mM NaCl, 1% Nonidet P-40, 1% sodium deoxycholate and 0.05% SDS).

2. Proteins were denatured by heating to 95 C in SDS-reducing buffer and were separated by electrophoresis on 12.5% SDS-polyacrylamide gels. The proteins were electrophoretically transferred to Hybond-ECL nitrocellulose membranes.

3. Membranes were probed with ab19247 diluted 1:1,000, and bound antibody was detected using an ECL Western blotting analysis system (Amersham Pharmacia Biotech). A band at approx 10 kDa is detected.

I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

Question 2

Tuesday 04-April-2006

Does this antibody detect both mono- and poly-ubiquitinated proteins?

ANSWER:

  

Thank you for your enquiry.

It has not been determined whether ab14372 detects mono- or poly-ubiquitinated proteins. It would quite likely recognize both.

Ab19247 should recognize ubiquinated proteins in Western blots regardless of how may ubiquitin chains are present.

I hope this information helps. Please let us know if you need anything else.

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