The nonclassical class I HLA-E molecule folds in the presence of peptide ligands donated by the signal sequences of permissive class I HLA alleles, with the aid of TAP and tapasin. To identify HLA-E-specific Abs, four monoclonals of the previously described MEM series were screened by isoelectric focusing (IEF) blot and immunoprecipitation/IEF on >30 single-allele class I transfectants and HLA-...

points and for determining the presence of charged mojeties on unknown antibiotics using minimal(microgram)quantities ofcrude material. Such infbrmatlon would also be valuable for identifying the presence of known substances in an antibiotic screen. However, IEF has not been apPhed extensively to antibiotics because their smaller size makes detection by traditional staining techniques difficult.

Isoelectric Focusing in Immobilized pH Gradient Strips Using the IPGphor Unit: Sample In-Gel Rehydration Angelika Görg, Oliver Drews, and Walter Weiss This protocol was adapted from "IEF in IPG Strips Using the IPGphor Unit," contributed by Angelika Görg, Oliver Drews, and Walter Weiss, Chapter 16, in Purifying Proteins for Proteomics (ed. Simpson). Cold Spring Harbor Laboratory Press, Cold Spr...

We review microfluidic devices designed for multidimensional sample analysis, with a primer on relevant theory, an emphasis on protein analysis, and an eye towards future improvements and challenges to the field. Image shows results of an on-chip IEF-CE separation of a protein mixture; unpublished surface plot data from A. E. Herr.

Methods Seventy fetuses at 19 to 28 weeks of gestation with an IEF and 63 controls were screened using two-dimensional and power Doppler echocardiography. DV wave velocities, DV velocity ratios, and diastolic time intervals were measured. The aortic peak velocity, pulmonary artery peak velocity, atrioventricular early-diastolic filling velocity (E), atrial contraction velocity (A), and E/A rati...

In a UK survey of the occurrence of extended spectrum beta-lactamases, 96 hospitals submitted a total of 3951 non-selected, non-duplicate isolates of Enterobacteriaceae from 100 patients in each hospital, 206 of these cultures being mixed and, therefore, discarded. These isolates were initially screened for strains likely to produce extended-spectrum beta-lactamases (ESBLs) by MIC determination...

The activity of sigma B, a secondary sigma factor of Bacillus subtilis, is primarily controlled by an anti-sigma factor protein (RsbW) that binds to sigma B and blocks its ability to form an RNA polymerase holoenzyme (E-sigma B). Inhibition of sigma B by RsbW is counteracted by RsbV, a protein that is essential for the activation of sigma B-dependent transcription. When crude B. subtilis extrac...

The serotype b-specific carbohydrate antigen (SbAg) of Actinobacillus actinomycetemcomitans Y4 is reported to be the O antigen of lipopolysaccharide, and the highest titers of serum antibody reactive with A. actinomycetemcomitans in early-onset periodontitis (EOP) patients bind SbAg. These high titers of serum antibody reactive with SbAg are associated with a lesser extent and severity of perio...

OBJECTIVE To investigate glycosylation changes associated with rheumatoid arthritis (RA) by determining whether there are beta1,4-galactosyltransferase (GTase) isoforms specific to or altered in the serum of patients with RA. Methods. Serum GTase isoform profiles were determined using isoelectric focusing (IEF) in patients with active RA (n = 9), disease controls (DC; n = 9), and healthy indivi...

An integrated protein concentration/separation system, combining non-native isoelectric focusing (IEF) with sodium dodecyl sulfate (SDS) gel electrophoresis on a polymer microfluidic chip, is reported. The system provides significant analyte concentration and extremely high resolving power for separated protein mixtures. The ability to introduce and isolate multiple separation media in a plasti...