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J Immunol Methods. 1986 Mar 13;87(2):283-7.

A simple method for the production of specific antiserum to the second nuclear antigen (EBNA-2) of Epstein-Barr virus (EBV).


A simple technique for raising specific antiserum to the native molecule of Epstein-Barr virus (EBV)-encoded nuclear antigen-2 (EBNA-2) from Raji cells is described. The procedure involves the use of immunoblotting to identify the EBNA-2 polypeptide followed by subsequent excision from an SDS-gel and immunization of experimental animals. The anti-EBNA-2 antiserum recognized a single polypeptide of 86-87 kDa on immunoblots prepared from extracts of EBV-positive Raji or B95-8 cells, while it did not react with any proteins form P3HR-1 or Daudi cells, which carry EBNA-2-defective virus. The method might be applicable to other systems where isolation of purified proteins for immunization is either difficult or unfeasible.

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