抗体抗原实验

Antibodies may be purified from serum, ascitic fluid, and tissue culture supernatant by a number of methods. Purified antibodies can be treated to produce fragments that may have enhanced properties for use in immunochemistry.

A major source of human antibodies are phage display libraries, which are constructed from various genetic sources. Antibodies are expressed as scFV and Fab antibody fragments using various vector systems. This review offers a comprehensive overview of M13 phage display antibody vec ...

Horseradish peroxidase is coupled to IgG antibody in a two-step procedure. In the first step monosaccharide residues in the enzyme are oxidized with periodate to produce aldehyde groups. Then, in the second step, the aldehyde groups are allowed to react with amino groups in the IgG antibody. The Sc ...

Alkaline phosphatase is coupled to immunoglobin G antibody in a one-step procedure using the homobifunctional reagent glutaraldehyde, which reacts with amino groups in the two proteins. The procedure is simple to perform and requires minimal equipment.

This valuable and unique book contains a compendium of methods and reviews that does much more than allow one to study the biology of neutrophils. What makes this collection of contributions so special is that it highlights and facilitates using the neutrophil as a simple, pure, single primary ce ...

Venous blood provides a ready source of large numbers of unstimulated granulocytes and mononuclear cells. Exploiting the differences in the relative densities of the leukocytes circulating in venous blood, one can separate leukocytes from erythrocytes as well as isolate the indivi ...

Advances in the understanding of neutrophil biochemistry require the development of effective procedures for isolating purified neutrophil populations. Although methods for human neutrophil isolation are now standard, similar procedures for isolating neutrophils fr ...

The neutrophil has long been recognized for its impressive number of cytoplasmic granules that harbor proteins indispensable for innate immunity. Analysis of isolated granules has provided important information on how the neutrophil grades its response to match the challenges it ...

The past decade has seen increasing use of the patch clamp technique on neutrophils and eosinophils. The main goal of these electrophysiological studies has been to elucidate the mechanisms underlying the phagocyte respiratory burst. NADPH oxidase activity, which defines the respi ...

Neutrophil-derived granule enzymes, oxidants, and mediators have been implicated in the pathogenesis of a variety of inflammatory diseases. Neutrophil apoptosis is associated with the loss of expression of adhesion molecules and greatly reduced responsiveness to external st ...

The detection of Ras superfamily GTPase activity in neutrophils is important when studying signaling events elicited by various ligands and cellular processes. Substantial progress in monitoring GTPase activation has been made in recent years by the development of high-affinity ...

Phospholipid metabolizing enzymes are important participants in neutrophil signal transduction pathways. The methods discussed herein describe assays for assessing the activities of phospholipase (PL)A2, PLC, PLD, and phosphoinositide 3-OH-kinase (PI3-K) in intact neu ...

Polymorphonuclear neutrophils play a key role in host defense and inflammation. Neutrophils can be activated by a variety of soluble and particulate factors, leading to an increase in the phosphorylation of numerous proteins on tyrosines, serines and threonines. Upon covalent bindi ...

Polymorphonuclear neutrophils (PMN) play an essential role in host defense against bacteria and fungi through coordinated responses such as adhesion, migration, phagocytosis, secretion, and activation of the NADPH oxidase. The mitogen-activated protein kinases (MAPKs) and t ...

Neutrophils and related phagocytic leukocytes are notoriously difficult to transfect, making the introduction of proteins into these cells for biological studies problematic. We describe here two methods that have been successfully used to introduce proteins into intact prim ...

Neutrophils are among the first cells to respond to acute inflammation through a multistep process initiated by selectin mediated rolling, which transitions to an integrin/intercellular adhesion molecule-dependent arrest and transmigration across endothelium. A confor ...

Neutrophil polarization and directed migration (chemotaxis) are critical for the inflammatory response. Neutrophil chemotaxis is achieved by the sensing of narrow gradients of chemoattractant and the subsequent polarization and directed migration toward the chemotact ...

We describe here a permeablization method that retains coupling between N-formylmethionyl- leucyl-phenylalanine (fMLP) receptor stimulation and barbed-end actin nucleation in neutrophils. Using fluorescently-tagged actin monomers, we are able to quantify and localize ...

Neutrophil migration from blood into tissues is required for effective innate immune responses against infection. Adhesion of the neutrophil in blood to the vascular endothelium and eventual migration through the vessel wall and accumulation at the site of infection involves diff ...

Neutrophils are short-lived granulocytes essential for innate host defense. We describe here methods for analysis of resting and activated cells using immunofluorescence and confocal microscopy. Procedures for stimulation of adherent and suspended cells are provided along ...