We have modified the columti-chromatographic technique of Mercer (1) to produce a sensitive and rapid separation of serum CK2 and CK3, even at normal total serum CK activity (3).12 However, CKj was not separated from CK2. Here, we report a modification of our method such that all three CK isoenzymes are separated by stepwise elution from a mini-column of diethylaminoethyl-cellulose. CK1 has onl...

1. Differentiation of the anodic (LDHi and LDH2) and cathodic (LDHs) isoenzymes of lactate dehydrogenase in human sera can be accomplished by combining variable substrate (lactate or pyruvate) concentrations with specific inhibitors such as urea. 2. One technic useful in the clinical laboratory is based upon a colorimetric assay for total LDH activity in which the oxidation of lactate is couple...

In an effort to clarify the issue of potentially false increases in creatine kinase (EC 2.7.3.2) MB isoenzyme (CK-MB) in uremia, we evaluated the CK profile of 84 persons undergoing chronic maintenance hemodialysis. We compared the performance of a new commercial two-site chemiluminometric immunoassay of CK-MB (Magic Lite; Ciba Corning Diagnostics) with that of electrophoresis on agarose gel (C...

1. The Michaelis constants for both isoenzymes for both substrates depend strongly on ionic concentration, being approximately proportional to phosphate concentration over considerable ranges. This is probably an effect of anions only. 2. In the absence of added salt, Km (2-oxoglutarate) (anionic isoenzyme) -is so small as to be .i.ndeterminate. 3. Km (L-aspartate) (anionic isoenzyme) passes th...

1. The Michaelis constants for both isoenzymes for both substrates depend strongly on ionic concentration, being approximately proportional to phosphate concentration over considerable ranges. This is probably an effect of anions only. 2. In the absence of added salt, K(m) (2-oxoglutarate) (anionic isoenzyme) is so small as to be indeterminate. 3. K(m) (l-aspartate) (anionic isoenzyme) passes t...

Lactate dehydrogenase C, an isoenzyme composed of C polypeptide subunits and found only in mature testes and spermatozoa, differs kinetically, chemically and immunologically from the five common isoenzymes of lactate dehydrogenase, each of which is a tetramer of A and/or B subunits. In the rat lactate dehydrogenase C exists in two molecular forms, isoenzymes C4 and A1C3. In addition to these tw...

Acetone fractionated peroxidase from crude extract of Brassica oleracea leaves (Cabbage) was purified in three steps on chromatographic columns, using Sp-Sepharose, DEAE-Sepharose and Con A-Sepharose. The specific activity of purified main isoenzyme (BOC-POD) is 1887 u/mg protein with RZ: 3.1, which is 172 times more than the RZ of crude extract with 4.3% recovery. The molecular weight of BOC-P...

Alpha-naphthyl acetate esterases (ANAE) were examined by cytochemical and isoelectric focusing (IEF) techniques in 48 cases of acute myeloid leukemia that were classified by conventional morphological criteria. Four main types of ANAE isoenzyme patterns were found by IEF, and comparisons with the expression of membrane receptors (Fc-lgG and C3b) and monocyte-specific antigens (UCHM1, UCHALF. an...

We have studied five patients who have exhibited an unusual alkaline phosphatase isoenzyme (ALP EC 3.1.3.1) migrating in an ultrafast position electrophoretically on cellulose acetate. This ALP isoenzyme has been identified in patients with benign and malignant liver diseases. In addition, a number of these patients exhibited a regular ALP liver isoenzyme and a fast (preliver) ALP isoenzyme in ...

Evidence of acute pancreatitis was sought in 35 patients with fulminant hepatic failure. Total amylase was raised in 22 patients and isoenzyme separation showed a distinct P3 isoenzyme (indicative of pancreatitis) in 14. In four patients with marked hyperamylasaemia (greater than 1000 U/l) the predominant isoenzyme was the salivary fraction. Pancreatic lipase was abnormally raised (greater than...