Non-denaturing gel electrophoresis system for the purification of membrane bound proteins

Journal Article · Fri Jan 01 00:00:00 EST 1988 · Prep. Biochem.; (United States)

DOI:https://doi.org/10.1080/00327488808062522· OSTI ID:6859780

Cavinato, A G; Macleod, R M; Ahmed, M S

A new method is described for the purification of a membrane bound glycoprotein, the kappa opioid receptor from human placental tissue. The method uses preparative slab-gel electrophoresis in the presence of the non-denaturing detergent CHAPS. A linear relationship between log molecular weight and SDS PAGE electrophoretic mobility of known molecular weight markers, in the presence of CHAPS, is observed. Using this method, we were able partially to purify an /sup 3/H-etorphine binding glycoprotein, from placental villus tissue, with an apparent molecular weight range of 60-70,000. The iodinated glycoprotein migrates in SDS PAGE with an apparent molecular weight of 63,000. This method may be useful for the isolation of membrane bound proteins, especially when an affinity ligand is not available.

• Research Organization: Memphis State Univ., TN (USA)
• OSTI ID: 6859780
• Journal Information: Prep. Biochem.; (United States), Vol. 18:2
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
GLUCOPROTEINS
FRACTIONATION
MEMBRANE PROTEINS
ELECTROPHORESIS
DETERGENTS
MOLECULAR WEIGHT
PLACENTA
PREGNANCY
TRACER TECHNIQUES
TRITIUM COMPOUNDS
ADDITIVES
CARBOHYDRATES
EMULSIFIERS
FETAL MEMBRANES
GLYCOPROTEINS
ISOTOPE APPLICATIONS
LABELLED COMPOUNDS
MEMBRANES
ORGANIC COMPOUNDS
PROTEINS
SACCHARIDES
SEPARATION PROCESSES
SURFACTANTS
WETTING AGENTS
550201* - Biochemistry- Tracer Techniques