platelet granules marker protein
j.broekman at verizon.net
Thu Dec 7 22:34:18 EST 2000
Dima Klenchin wrote:
> behrends at plexus.uke.uni-hamburg.de wrote:
> :Dear netters,
> :we do biochemical fractionation of human platelets
> :to seperate plasma membrane from intracellular
> :granules. We would need a good antibody for a protein
> :that is expressed in platelet granules to characterize
> :our fractions that we get over a sucrose gradient.
> :So far we have found CD63 and a polyclonal and a mono-
> :clonal antibody is available from Santa Cruz. But there
> :are as yet no publications with Western blots and we
> :do not know whether CD63 expression in human platelets
> :granules is high enough to be detected by these antibodies.
> :I imagine that there should be other proteins in the
> :membrane of human platelet granules that are better
> :characterized and where antibodies are procurable for
> :Western blot.
> Platelets secrete hexosaminidase and platelet factor 4
> through granule exocytosis. There should be Ab for both
> available. Additionally, it is said that Rab6 localizes almost
> exclusively to platelet dense core granules. Anti Rab6
> are available for sure.
> - Dima
Fibrinogen is an alpha granule protein that is secreted along with many
coagulation factors when platelets are stimulated. If you manage to
disrupt platelets without activating them, you should be able to find
many proteins that can perform for your purpose. My own work from some
25 years ago might provide one of many starting points for you:
M. J. Broekman, N. P. Westmoreland, and P. Cohen. An improved method for
isolating alpha granules and mitochondria from human platelets. J.Cell
Biol. 60:507-519, 1974.
M. J. Broekman, R. I. Handin, and P. Cohen. Distribution of fibrinogen,
and platelet factors 4 and XIII in subcellular fractions of human
platelets. Br.J.Haematol. 31:51-55, 1975.
M. J. Broekman, R. I. Handin, A. Derksen, and P. Cohen. Distribution of
phospholipids, fatty acids, and platelet factor 3 activity among
subcellular fractions of human platelets. Blood 47:963-971, 1976.
"Later" papers (the Kaplan paper was an early Citation Classic):
K. L. Kaplan, M. J. Broekman, A. Chernoff, G. R. Lesznik, and M.
Drillings. Platelet alpha-granule proteins: studies on release and
subcellular localization. Blood 53:604-618, 1979.
M. B. Zucker, M. W. Mosesson, M. J. Broekman, and K. L. Kaplan. Release
of platelet fibronectin (cold-insoluble globulin) from alpha granules
induced by thrombin or collagen; lack of requirement for plasma
fibronectin in ADP-induced platelet aggregation. Blood 54:8-12, 1979.
(Please answer to the newsgroup only, I will not answer email)
More information about the Methods