TY - JOUR
T1 - Native species of helix destabilizing protein-1 in mouse myeloma identified by antibody probing of Western blots
AU - Planck, Stephen R.
AU - Wilson, Samuel H.
N1 - Funding Information:
Laboratory of Biochemistry, National Cancer Institute National Institutes of Health, Bethesda, MD 20205 +Department of Anatomy, Oregon Health Sciences University Portland, Oregon 97201
PY - 1985/8/30
Y1 - 1985/8/30
N2 - Antibody probing of Western blots is a method for analyzing the apparent Mr of a protein in any given preparation (Renart, J., Reiser, J., and Stark, G., Proc. Natl. Acad. Sci. USA 76: 3116, 1979). We prepared a rabbit antiserum to purified mouse myeloma helix destabilizing protein-1 and used this antiserum in Western blotting experiments with a crude homogenate of mouse myeloma. The results indicated that the native species of helix destabilizing protein-1 can be degraded during purification. This in vitro proteolysis results in complete loss of the native species and accumulation of lower Mr proteins that represent limit digestion products. These findings have identified the true native form of mouse myeloma HDP-1 as a protein of apparent Mr=36,000 to 38,000, instead of the Mr=24,000 and 27,000 proteins obtained by routine purification.
AB - Antibody probing of Western blots is a method for analyzing the apparent Mr of a protein in any given preparation (Renart, J., Reiser, J., and Stark, G., Proc. Natl. Acad. Sci. USA 76: 3116, 1979). We prepared a rabbit antiserum to purified mouse myeloma helix destabilizing protein-1 and used this antiserum in Western blotting experiments with a crude homogenate of mouse myeloma. The results indicated that the native species of helix destabilizing protein-1 can be degraded during purification. This in vitro proteolysis results in complete loss of the native species and accumulation of lower Mr proteins that represent limit digestion products. These findings have identified the true native form of mouse myeloma HDP-1 as a protein of apparent Mr=36,000 to 38,000, instead of the Mr=24,000 and 27,000 proteins obtained by routine purification.
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U2 - 10.1016/0006-291X(85)91811-X
DO - 10.1016/0006-291X(85)91811-X
M3 - Article
C2 - 2994656
AN - SCOPUS:0022235502
SN - 0006-291X
VL - 131
SP - 362
EP - 369
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
IS - 1
ER -