immunoperoxidase labeling was then graded for intensity and examined for artifacts. One monoclonal antibody against MBP and one against NF resulted in labeling that was It has been demonstrated in the past that variations in fixatives and tissue processing techniques can affect the results of immunological staining obtained by antisera (3,30). These variations may be due to antigen loss, alteration, or inacces-'Supported in part by grants from the National Multiple Sclerosis Society (RC1 l61-A-3

No abstract

The metabolism of arachidonic acid into inflammatory mediators (e.g. prostaglandin, leukotrienes) is dependent upon the rate-limiting enzyme phospholipase A(2). Localization and quantification of type II 14 kDa phospholipase A(2) (PLA(2)) in cells or tissue preparations has historically been accomplished through activity measurements, a process that can provide variable results due to interference by exogenous substances with hydrolysis assessment. Others have reported on the use of sandwich enzyme immunoassays (EIA) to measure 14 kDa PLA(2) by mass in serum and exudate fluids, e.g. synovial fluid. Herein, we report the utilization of a human recombinant type II 14 kDa PLA(2) sandwich EIA to directly measure cell or tissue-residing 14 kDa PLA(2). It is known that type II 14 kDa PLA(2) resists acid treatment, and this technique was applied to cell fractions which liberated the enzyme from cellular membrane components prior to quantitation by EIA. Two human immune cell populations were assessed and shown to contain measurable levels of 14 kDa PLA(2). Neutrophil or monocyte cytosolic fractions contained no measurable levels whereas the respective 100 000g particulate fractions contained 2.6+/-0.8 pg (neutrophil) and 2.1+/-0.6 pg (monocyte) 14 kDa PLA(2)/mug protein. Human placenta cytosolic fractions contained no measurable levels while 100 000g particulate contained approximately 25 ng 14 kDa PLA(2)/mg protein. This EIA, in conjunction with acid extraction, provides an easy and reproducible assay to identify and quantify this enzyme in cells and whole tissues, expanding our ability to study the relationship of this enzyme to inflammatory processes.

No abstract

We describe here a novel IgG monoclonal antibody to erythroid-related factor (ERAF), also known as alpha hemoglobin stabilizing protein (AHSP) and eryththroid differentiation related factor (EDRF). Our antibody named PCE 5 is an IgG(1) kappa chain and is to the peptide sequence MVTVVE ranked highly in our active site analysis and binds with high affinity to ERAF.