Radioactive labeling of proteins in cultured postimplantation mouse embryos. II. Dose and time dependency

Journal Article · Sat Jul 01 00:00:00 EDT 1989 · In Vitro Cellular and Developmental Biology; (USA)

DOI:https://doi.org/10.1007/BF02623634· OSTI ID:5673535

Nowak, J; Klose, J ^[1]

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1. Freie Universitaet, Berlin (Germany, F.R.)

The conditions for optimum incorporation of radioactive amino acids into proteins of cultured postimplantation mouse embryos were investigated under the aspect of using these proteins for two-dimensional electrophoretic separations and fluorography. The aim was to obtain highly radioactively labeled proteins under conditions as physiological as possible. Mouse embryos of Days 8, 10, and 11 of gestation were cultured in Tyrode's solution. Incubation time and concentration of ({sup 3}H (or {sup 14}C))amino acids in the culture medium were varied over a broad range. Embryos were prepared with placenta and yolk sac or without any embryonic envelopes. After culturing, the physiologic-morphologic state of the embryos was registered on the basis of several criteria. The radioactivity taken up by the total protein of each embryo was determined and calculated in disintegrations per minute per milligram protein per embryo. To approach our aim, embryos of different developmental stages had to be cultured under different conditions. A good compromise for Day-8, Day-10, and Day-11 embryos was: embryos prepared with yolk sac (opened) and placenta, 150 microCi radioactive amino acids added per milliliter medium, incubation for 4 to 5 h. For maximum labeling of proteins it is advisable to culture Day-10 embryos without embryonic envelopes under particular conditions.

• OSTI ID: 5673535
• Journal Information: In Vitro Cellular and Developmental Biology; (USA), Vol. 25:7; ISSN 0883-8364
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
PROTEINS
LABELLING
AMINO ACIDS
CARBON 14 COMPOUNDS
CULTURE MEDIA
EMBRYOS
MICE
ONTOGENESIS
PLACENTA
PREGNANCY
SAMPLE PREPARATION
TIME DEPENDENCE
TISSUE CULTURES
TRACER TECHNIQUES
TRITIUM COMPOUNDS
TWO-DIMENSIONAL ELECTROPHORESIS
ANIMALS
CARBOXYLIC ACIDS
ELECTROPHORESIS
FETAL MEMBRANES
HYDROGEN COMPOUNDS
ISOTOPE APPLICATIONS
LABELLED COMPOUNDS
MAMMALS
MEMBRANES
ORGANIC ACIDS
ORGANIC COMPOUNDS
RODENTS
VERTEBRATES
550201* - Biochemistry- Tracer Techniques