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We have raised a monoclonal antibody (mAb) (HFD9) that detects a 28 kDa protein (p28) enriched in the Golgi membrane. p28 was localized to the perinuclear Golgi region in all cell lines thus far examined. Its Golgi localization was confirmed by its colocalization with Golgi markers using indirect immunofluorescence microscopy. Immunogold labelling demonstrates that the majority of p28 was localized on the cis-Golgi and its associated structures. Two independent experiments demonstrate that the p28 epitope recognized by mAb HFD9 is exposed to the cytosol. Extraction of Golgi membranes with a variety of reagents revealed that p28 behaves like an integral membrane protein. mAb HFD9 thus defines a novel 28 kDa integral membrane protein on the cis-Golgi. To our knowledge, p28 represents the first integral membrane protein of the Golgi system identified via the antibody approach whose epitope is cytoplasmically-oriented and highly-conserved. Monoclonal antibody HFD9 will thus provide a useful tool for further studies on the cis side of the Golgi, which is not well characterised due to the lack of good markers.
V N Subramaniam, and
J Krijnse-Locker, and
B L Tang, and
M Ericsson, and
A R Yusoff, and
G Griffiths, and
W Hong
August 1992,
Cell structure and function,
V N Subramaniam, and
J Krijnse-Locker, and
B L Tang, and
M Ericsson, and
A R Yusoff, and
G Griffiths, and
W Hong
July 1998,
FEBS letters,
V N Subramaniam, and
J Krijnse-Locker, and
B L Tang, and
M Ericsson, and
A R Yusoff, and
G Griffiths, and
W Hong
January 1982,
The EMBO journal,
V N Subramaniam, and
J Krijnse-Locker, and
B L Tang, and
M Ericsson, and
A R Yusoff, and
G Griffiths, and
W Hong
February 1983,
Archives of biochemistry and biophysics,
V N Subramaniam, and
J Krijnse-Locker, and
B L Tang, and
M Ericsson, and
A R Yusoff, and
G Griffiths, and
W Hong
January 1995,
Molecular membrane biology,
V N Subramaniam, and
J Krijnse-Locker, and
B L Tang, and
M Ericsson, and
A R Yusoff, and
G Griffiths, and
W Hong
September 1997,
Clinical and diagnostic laboratory immunology,
V N Subramaniam, and
J Krijnse-Locker, and
B L Tang, and
M Ericsson, and
A R Yusoff, and
G Griffiths, and
W Hong
May 1995,
Histochemistry and cell biology,
V N Subramaniam, and
J Krijnse-Locker, and
B L Tang, and
M Ericsson, and
A R Yusoff, and
G Griffiths, and
W Hong
September 1985,
Molecular and biochemical parasitology,
V N Subramaniam, and
J Krijnse-Locker, and
B L Tang, and
M Ericsson, and
A R Yusoff, and
G Griffiths, and
W Hong
December 1994,
Journal of cell science,
V N Subramaniam, and
J Krijnse-Locker, and
B L Tang, and
M Ericsson, and
A R Yusoff, and
G Griffiths, and
W Hong
January 1993,
FEBS letters,
