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Production and characterization of antipeptide antibodies against human O6-methylguanine-DNA methyltransferase.

Publication ,  Journal Article
Ostrowski, LE; Pegram, CN; Von Wronski, MA; Humphrey, PA; He, XM; Shiota, S; Mitra, S; Brent, TP; Bigner, DD
Published in: Cancer Res
July 1, 1991

Four synthetic peptides from the sequence of human O6-methylguanine-DNA methyltransferase (MGMT), three corresponding to different hydrophilic regions and one corresponding to the sequence containing the alkyl acceptor residue cysteine 145, were used to immunize rabbits. The antibody against Peptide III (residues 171-184) was highly specific, and MGMT protein could be detected on Western blots of soluble protein extracts containing as little as 1 fmol of active MGMT. Antibodies against all of the peptides were able to immunoprecipitate denatured MGMT, while only the antibody against Peptide III was able to react with active enzyme. The antibody against Peptide III did not cross-react with methyltransferase from mice. The use of synthetic peptides has led to the production of a highly sensitive, specific antibody that recognizes native and denatured human MGMT. This antibody should prove useful in studies involving the detection, purification, and characterization of this enzyme.

Duke Scholars

Published In

Cancer Res

ISSN

0008-5472

Publication Date

July 1, 1991

Volume

51

Issue

13

Start / End Page

3339 / 3344

Location

United States

Related Subject Headings

  • Species Specificity
  • Solubility
  • Precipitin Tests
  • Peptides
  • Oncology & Carcinogenesis
  • O(6)-Methylguanine-DNA Methyltransferase
  • Molecular Sequence Data
  • Methyltransferases
  • Humans
  • Blotting, Western
 

Citation

APA
Chicago
ICMJE
MLA
NLM
Ostrowski, L. E., Pegram, C. N., Von Wronski, M. A., Humphrey, P. A., He, X. M., Shiota, S., … Bigner, D. D. (1991). Production and characterization of antipeptide antibodies against human O6-methylguanine-DNA methyltransferase. Cancer Res, 51(13), 3339–3344.
Ostrowski, L. E., C. N. Pegram, M. A. Von Wronski, P. A. Humphrey, X. M. He, S. Shiota, S. Mitra, T. P. Brent, and D. D. Bigner. “Production and characterization of antipeptide antibodies against human O6-methylguanine-DNA methyltransferase.Cancer Res 51, no. 13 (July 1, 1991): 3339–44.
Ostrowski LE, Pegram CN, Von Wronski MA, Humphrey PA, He XM, Shiota S, et al. Production and characterization of antipeptide antibodies against human O6-methylguanine-DNA methyltransferase. Cancer Res. 1991 Jul 1;51(13):3339–44.
Ostrowski, L. E., et al. “Production and characterization of antipeptide antibodies against human O6-methylguanine-DNA methyltransferase.Cancer Res, vol. 51, no. 13, July 1991, pp. 3339–44.
Ostrowski LE, Pegram CN, Von Wronski MA, Humphrey PA, He XM, Shiota S, Mitra S, Brent TP, Bigner DD. Production and characterization of antipeptide antibodies against human O6-methylguanine-DNA methyltransferase. Cancer Res. 1991 Jul 1;51(13):3339–3344.

Published In

Cancer Res

ISSN

0008-5472

Publication Date

July 1, 1991

Volume

51

Issue

13

Start / End Page

3339 / 3344

Location

United States

Related Subject Headings

  • Species Specificity
  • Solubility
  • Precipitin Tests
  • Peptides
  • Oncology & Carcinogenesis
  • O(6)-Methylguanine-DNA Methyltransferase
  • Molecular Sequence Data
  • Methyltransferases
  • Humans
  • Blotting, Western