Abstract
This paper describes the isolation of plasma membrane vesicles formed by nitrogen cavitation of canine neutrophils. Plasma membranes from disrupted cells were separated from other membranes and organelles by Percoll-density gradient centrifugation. Transmission electron microscopic examination of membrane preparations chromatographed on either Sephacryl S-1000 or Sepharose 4B revealed that two populations of plasma membrane vesicles were formed: large (176 ± 22 nm), and small (119 ± 11 nm). Purified large vesicles were separated from Percoll and contaminating cytosol by Sephacryl S-1000 chromatography. Small vesicles were obtained free of Percoll by recavitating purified large vesicles. Problems encountered due to the presence of a soluble 5'-nucleotidase inhibitor also are discussed. Large and small membrane vesicles were separated into adherent and non-adherent populations by affinity chromatography on either concanavalin A-sepharose or lentil lectin-Sepharose columns.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 353-368 |
| Number of pages | 16 |
| Journal | Journal of Leukocyte Biology |
| Volume | 38 |
| Issue number | 3 |
| State | Published - 1985 |
| Externally published | Yes |
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ASJC Scopus subject areas
- Cell Biology
Cite this
Isolation of canine neutrophil plasma membranes. / O'Donnell, Robert T; Andersen, B. R.
In: Journal of Leukocyte Biology, Vol. 38, No. 3, 1985, p. 353-368.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Isolation of canine neutrophil plasma membranes
AU - O'Donnell, Robert T
AU - Andersen, B. R.
PY - 1985
Y1 - 1985
N2 - This paper describes the isolation of plasma membrane vesicles formed by nitrogen cavitation of canine neutrophils. Plasma membranes from disrupted cells were separated from other membranes and organelles by Percoll-density gradient centrifugation. Transmission electron microscopic examination of membrane preparations chromatographed on either Sephacryl S-1000 or Sepharose 4B revealed that two populations of plasma membrane vesicles were formed: large (176 ± 22 nm), and small (119 ± 11 nm). Purified large vesicles were separated from Percoll and contaminating cytosol by Sephacryl S-1000 chromatography. Small vesicles were obtained free of Percoll by recavitating purified large vesicles. Problems encountered due to the presence of a soluble 5'-nucleotidase inhibitor also are discussed. Large and small membrane vesicles were separated into adherent and non-adherent populations by affinity chromatography on either concanavalin A-sepharose or lentil lectin-Sepharose columns.
AB - This paper describes the isolation of plasma membrane vesicles formed by nitrogen cavitation of canine neutrophils. Plasma membranes from disrupted cells were separated from other membranes and organelles by Percoll-density gradient centrifugation. Transmission electron microscopic examination of membrane preparations chromatographed on either Sephacryl S-1000 or Sepharose 4B revealed that two populations of plasma membrane vesicles were formed: large (176 ± 22 nm), and small (119 ± 11 nm). Purified large vesicles were separated from Percoll and contaminating cytosol by Sephacryl S-1000 chromatography. Small vesicles were obtained free of Percoll by recavitating purified large vesicles. Problems encountered due to the presence of a soluble 5'-nucleotidase inhibitor also are discussed. Large and small membrane vesicles were separated into adherent and non-adherent populations by affinity chromatography on either concanavalin A-sepharose or lentil lectin-Sepharose columns.
UR - http://www.scopus.com/inward/record.url?scp=0021822909&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0021822909&partnerID=8YFLogxK
M3 - Article
C2 - 2993461
AN - SCOPUS:0021822909
VL - 38
SP - 353
EP - 368
JO - Journal of Leukocyte Biology
JF - Journal of Leukocyte Biology
SN - 0741-5400
IS - 3
ER -