A modified procedure for the determination of loukocyte alkaline phosphatase
TL;DR: A satisfactory assay for the leukocyte enzyme employing 2-amino-2-methyl-1-propanol buffer is outlined, and the reaction is not linear with time when run in this buffer.
About: This article is published in Biochemical Medicine.The article was published on 1970-08-01. It has received 161 citations till now. The article focuses on the topics: Alkaline phosphatase & Glycine.
Citations
More filters
TL;DR: Data indicate that the b- Cytochrome is in the membrane of the specific granules of unstimulated neutrophils and that stimulus-induced fusion of these granules with the plasma membrane results in a translocation of the cytochrome.
Abstract: We describe a new method for subcellular fractionation of human neutrophils. Neutrophils were disrupted by nitrogen cavitation and the nuclei removed by centrifugation. The postnuclear supernatant was applied on top of a discontinuous Percoll density gradient. Centrifugation for 15 min at 48,000 g resulted in complete separation of plasma membranes, azurophil granules, and specific granules. As determined by ultrastructure and the distribution of biochemical markers of these organelles, approximately 90% of the b-cytochrome in unstimulated cells was recovered from the band containing the specific granules and was shown to be in or tightly associated with the membrane. During stimulation of intact neutrophils with phorbol myristate acetate or the ionophore A23187, we observed translocation of 40-75% of the b-cytochrome to the plasma membrane. The extent of this translocation closely paralleled release of the specific granule marker, vitamin B12-binding protein. These data indicate that the b-cytochrome is in the membrane of the specific granules of unstimulated neutrophils and that stimulus-induced fusion of these granules with the plasma membrane results in a translocation of the cytochrome. Our observations provide a basis for the assembly of the microbicidal oxidase of the human neutrophil.
906 citations
TL;DR: The aim of the present study was to determine the subcellular localization of NGAL and an enzyme-linked immunosorbent assay for NGAL was developed using specific anti-NGAL antibodies, which confirmed the existence of differentially mobilized granule subpopulations among peroxidase negative granules.
372 citations
TL;DR: HCAP-18 as discussed by the authors is the only human member of the antibacterial and endotoxin-binding family of proteins known as cathelicidins, and it was found to colocalize and comobilize with lactoferrin, but not with gelatinase in subcellular fractions.
339 citations
TL;DR: Results suggest that PMA activates the cell in the same manner as does phagocytosis, and this compound should provide a useful tool for elucidating the metabolic events underlying the phenomena of phagcytosis and bacterial killing by polymorphonuclear leukocytes.
335 citations
TL;DR: A 19 kDa protein was identified in specific granules of human neutrophils, stored as a ‘pro‐peptide’ in specificgranules, based on amino‐acid sequences of isolated tryptic fragments that were in complete agreement with the sequence deduced from the cDNA.
334 citations
References
More filters
TL;DR: An investigation of the biochemical changes following experimental liver injury felt the need of a simple, rapid, and accurate method for determining the protein fractions in small amounts of serum and began with Kingsley’s biuret procedure.
15,717 citations
1,045 citations
Journal Article•
198 citations
TL;DR: The leukocytes were investigated and it was found that on incubation in serum or saline the cell alkaline phosphatase activity increased, however, this in vitro rise was unrelated to the source of the serum menstrum.
106 citations
99 citations