|
|
|
|
 |
|||
|
|||
Immunoelectron microscopic localization of phosphoproteins associated with the mitotic spindle
DD Vandre and RW Burry
Department of Cell Biology, Neurobiology, and Anatomy, Ohio State University, Columbus 43210-1239.
We examined the immunogold staining of microtubules and microtubule organizing centers using an improved silver-enhancement reagent for small (1-1.4 nm) gold-conjugated secondary antibodies. First, the staining properties of different commercial preparations of gold- labeled antibodies were compared for sample penetration, label uniformity, and labeling density, and Nanogold 1.4-nm gold-conjugated F(ab') was found to be superior to the other probes examined. However, in samples examined for the localization of alpha- and beta-tubulin, gold staining did not extend through the pericentriolar material nor were the centrioles labeled. This apparent lack of centrosomal staining was not due to problems associated with penetration of the antibody probes, since staining adjacent to and within the centriolar cylinder was observed when phosphoprotein antigens recognized by the MPM-2 antibody were localized. The MPM-2 antibodies also localized to mitotic kinetochores, kinetochore fibers, and midbodies, in addition to mitotic centrosomes. The level of MPM-2 staining of the centrosome varied through the cell cycle. At interphase, this staining was restricted within the centriolar cylinder, whereas in mitotic cells extensive staining throughout the pericentriolar material was also observed. These results established the close relationship of MPM-2-reactive phosphoproteins with the centrosome, and suggest that this technique may be useful for ultrastructural localization of other cytoskeletal proteins.
Volume 40,
Issue 12,
pp. 1837-1847,
12/01/1992
Copyright © 1992 by The Histochemical Society
CiteULike 
Complore 
Connotea 
Del.icio.us 
Digg 
Reddit 
Technorati What's this?
This article has been cited by other articles:
|
|
 |
|
  N. Ma, S. Matsunaga, H. Takata, R. Ono-Maniwa, S. Uchiyama, and K. Fukui Nucleolin functions in nucleolus formation and chromosome congression J. Cell Sci., June 15, 2007; 120(12): 2091 - 2105. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. F. Hainfeld and R. D. Powell New Frontiers in Gold Labeling J. Histochem. Cytochem., April 1, 2000; 48(4): 471 - 480. [Abstract] [Full Text]
|
|
|
|
|
|
J. M. Robinson, T. Takizawa, and D. D. Vandré Enhanced Labeling Efficiency Using Ultrasmall Immunogold Probes: Immunocytochemistry J. Histochem. Cytochem., April 1, 2000; 48(4): 487 - 492. [Abstract] [Full Text]
|
|
|
|
 |
|
 P. A. Wigge, O. N. Jensen, S. Holmes, S. Soues, M. Mann, and J. V. Kilmartin Analysis of the Saccharomyces Spindle Pole by Matrix-assisted Laser Desorption/Ionization (MALDI) Mass Spectrometry J. Cell Biol., May 18, 1998; 141(4): 967 - 977. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S Soues and I. Adams SPC72: a spindle pole component required for spindle orientation in the yeast Saccharomyces cerevisiae J. Cell Sci., January 9, 1998; 111(18): 2809 - 2818. [Abstract] [PDF]
|
|
|
|
|
|
C Roghi, R Giet, R Uzbekov, N Morin, I Chartrain, R Le Guellec, A Couturier, M Doree, M Philippe, and C Prigent The Xenopus protein kinase pEg2 associates with the centrosome in a cell cycle-dependent manner, binds to the spindle microtubules and is involved in bipolar mitotic spindle assembly J. Cell Sci., January 3, 1998; 111(5): 557 - 572. [Abstract] [PDF]
|
|
|
|
|
|
E. Paul and A Quaroni Identification of a 102 kDa protein (cytocentrin) immunologically related to keratin 19, which is a cytoplasmically derived component of the mitotic spindle pole J. Cell Sci., January 11, 1993; 106(3): 967 - 981. [Abstract] [PDF]
|
|
| Guidelines | Subscriptions | About | exPRESS - Current - Archive | Business Information | Contact |