This report describes the development of a new panel of monoclonal antibodies established after immunization of mice with purified surfactant protein D of the rat. To enhance the detection of SP-D in formalin- or Schaffer-fixed samples, immunohistochemistry was performed by using microwave pretreatment of paraffin sections. Using these new antibodies that bind to type II epithelial cells, Clara cells, and alveolar macrophages, the responses of lung parenchymal cells were examined in a radiation-induced fibrosis model. Increased accumulation of extracellular SP-D in the alveolar space was found. Double staining with anti-surfactant protein A antibodies revealed different Clara cell populations containing one or both types of surfactant proteins.

Animals↗

PubMedDate not supplied

[The acetylcholinesterase of Bungarus multicinctus venom. Purification and properties (author's transl)].

Acetylcholinesterase from Banded krait (Bungarus multicinctus) venom has been purified by CM-Sephadex chromatography and affinity chromatography to a specific activity of 4290 U/mg. The purified enzyme is a glycoprotein. It is free of electrophoretically detectable contaminating proteins. A molecular weight of 140,000 +/- 5,000 has been determined by gradient gel electrophoresis for the native enzyme. It is split into two equal-sized subunits (Mr 70,000 +/- 2,000) by SDS treatment. The N-terminal amino acid analysis gave glycine and serine. The purified acetylcholinesterase can be resolved by disc gel electrophoresis into four and by isoelectric focusing into six isozymes. The pI value of the main isozyme has been found to be 5.98 +/- 0.05.

Acetylcholinesterase↗

PubMedDate not supplied

[Acetylcholinesterase from bovine erythrocytes. Purification and properties of the enzyme solubilized in the presence and the absence of Triton X-100 (author's transl)].

Acetylcholinesterase was released from bovine erythrocytes by Triton X-100 treatment and purified by twwfold affinity chromatography. The detergentfree enzyme was obtained with a specific activity of 4130 U/mg (303 000-fold purification) and a 25% yield. Alternatively, the commercial available crude enzyme was purified. The latter preparation has an uniform molecular weight (Mr 175 000). The Triton-solubilized enzyme, however, can be resolved after removal of the detergent in eight multiple forms (Mr 175 000 and multiple values), in the presence of Triton there exists only one form (Mr 338 000). The amino acid composition of the two enzyme preparations differs significantly. No differences were observed with respect to other properties: SDS gel electrophoresis revealed two protein bands (Mr 166 000 and 86 000) with both preparations. The enzyme is a glycoprotein with a pI value of 4.3 and contains strongly bound phosphatidylethanolamine. The N-terminal amino acid has been found to be Glu (or Gln).

Acetylcholinesterase↗