Purification and measurement of calpromotin, the cytoplasmic protein which activates calcium-dependent potassium transport
| Autor: | S K Shriver, R B Moore, Gordon A. Plishker |
|---|---|
| Rok vydání: | 1990 |
| Předmět: |
Erythrocytes
Potassium Biophysics chemistry.chemical_element Biological Transport Active Enzyme-Linked Immunosorbent Assay Calcium Biochemistry Chromatography DEAE-Cellulose chemistry.chemical_compound Column chromatography Potassium phosphate Reference Values medicine Humans Molecular Biology Polyacrylamide gel electrophoresis Ion transporter Gel electrophoresis Chromatography Chemistry Cell Biology Blood Proteins Molecular Weight Red blood cell medicine.anatomical_structure Chromatography Gel Electrophoresis Polyacrylamide Gel Carrier Proteins |
| Zdroj: | Biochemical and biophysical research communications. 166(1) |
| ISSN: | 0006-291X |
| Popis: | A simple procedure is described for the purification of calpromotin, a protein from the cytoplasm of red blood cells which is capable of activating calcium-dependent potassium transport. The purification steps involve a salt gradient elution from an anion exchange column (Whatman DE-52) followed by a potassium phosphate gradient elution from a column of hydroxyapatite (HA Ultrogel). These steps result in a 54% yield with a 161 fold purification. The calpromotin is estimated to be 99% pure as determined by densitometry of the protein profile on an SDS polyacrylamide gel. A competitive enzymelinked immunosorbent assay (ELISA) using rabbit anti-human calpromotin antibodies, is described for measuring levels of calpromotin in the 5 to 100 ng range. |
| Databáze: | OpenAIRE |
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